Guide for Collection

Biological Materials in
Private Hospitals

Version IX, 2010

Autor: Enfª Priscila Rosseto de Toledo. Data: 15/02/2010

Approved by: Dr. Elisa Junko Ura. Date: 03/22/2010. CONTROLLED

OBJECTIVE

Ensure the execution of collection procedures in accordance with the

existing standardization in the SIG, as well as the quality of the sample and

mainly the safety of the patient.

INDEX

CHAPTER I: LABORATORY PROCESS

CHAPTER II: THE PRE-ANALYTICAL PHASE

CHAPTER III: BLOOD COLLECTION

CHAPTER IV: MICROBIOLOGY

CHAPTER V: MISCELLANEOUS COLLECTIONS

THEORETICAL REFERENCE

ANNEXES
 CHAPTER I: LABORATORY PROCESS

The process of conducting laboratory tests begins with the requesting physician,
when filling out a medical request for laboratory tests and
it ends when this professional receives and interprets the obtained results. Among these
two points, the laboratory must register the patient and the exams
requested, the collection of samples, the transportation to the laboratory, the preparation of the

samples, the execution of the tests, the analysis of the obtained results, the release of the
results, the delivery of reports and assistance in interpreting the results. This
the process can be divided into 3 main phases:

PRE - ANALYTICAL PHASE

Medical Request
Patient and exam data registration in the computerized system
Identification of the prerequisites necessary for conducting the tests
Identification of medications in use
Collection of necessary materials
Identification of samples
Storage of samples for transport
Forwarding of samples to the laboratory
Preparation of samples for analysis

ANALYTICAL PHASE
Calibration of equipment
Preventive maintenance of equipment
Preparation of the necessary reagents
Conducting the exams

POST-ANALYTICAL PHASE
Analysis of the results obtained (including the analysis of the evolution of the results of
patient)
Release of the results
Delivery of reports
Support in the interpretation of results
The Quality Control programs used in the laboratory have a focus
important in controlling the precision and accuracy of the tests.
 CHAPTER II: THE PRE-ANALYTICAL PHASE

Definition

It is the phase that involves customer service regarding:

Medical requests;
Information provided for home collections;
Information about exam preparation;
Registration in the computerized system;
Identification of prerequisites and medications in use;
Collection of biological material;
Sample storage;
Transport to the support laboratory;
Preparation and/or screening of the sample for analysis.

Pre-analytical Errors

Pre-analytical errors are the mistakes made during the period before the sample analysis.
which can influence the quality of the final measured results and compromise the
diagnosis and treatment of the patient.

Basic Procedures to Minimize Error Occurrences

According to SBPC 2010, the collector must ensure that the sample will be collected from
patient specified in the examination request.
For adult and conscious patients
Request to provide full name, identity number, or date of birth.
• Compare this information with the constants in the exam request.
For hospitalized patients
In general, hospitals provide pre-printed labels with the information of
necessary identification. Even so, the collector must verify the identification in
bracelet or the identification posted at the entrance of the room, when available. The
The bed number should never be used as a criterion for identification. In units
closed, such as Intensive Care Units or Intermediate Units, the collector
should, in case of doubts in identification, seek help from the professionals of that
sector with the purpose of ensuring the proper identification of the patient.
Report any discrepancies in information to the laboratory supervisor.
For very young patients or with some type of communication difficulty
The collector must make use of information from an accompanying person or from
nursing.
Patients attended in the emergency room or in emergency rooms can be
identified by their name and entry number in the unit registration
emergency.

It is essential that the identification can be traced at any moment.

process.
The collected material must be identified in the presence of the patient. In the systems
manuals, this can be done by placing labels in the collection tubes with the
patient's name, the date of collection, and the sequential service number. This
number must appear on all documents, samples, work maps, reports
and the final report.

Pre-analytical Variables

They are variables that can alter the baseline/normal values of individuals, they are:

Chronobiological;
Gender;
Age;
Customer's position at the time of collection;
Physical activity;
Altitude (geographical location);
Trauma or pain;
Hyperthermia;
Emotional state;
Prolonged fasting;
Prolonged garrotting.

In hospital environments we have:

Therapeutic or diagnostic procedures (Blood transfusion; Infusion of

solutions among others).
Customer approach to the collection procedure.

The procedure for collecting biological material always causes insecurity for the client,
pois a espera deste resultado pode trazer consigo a idéia de alívio, medo, novos
diagnostic procedures, etc., for this reason it is important that at this moment the
The collector should be safe and confident so that they can reassure the customer.

If the patient does not exhibit changes in the level of consciousness, it is important that the

the collection procedure should be explained in clear language, even in case of

children. This facilitates the patient's collaboration in the procedure. When the same
not fit to understand the information, these are passed on to the companion
if this is present.

Never say 'it won't hurt at all', as the patient will lose trust if you come.
to perform another procedure.

If the patient asks "what is a specific exam for", say the name of the
exam, and that only the doctor or nurse will be able to explain the purpose of
test.

Efficiency and description are extremely important

It is vital that the professional is appropriately dressed, for this reason it is

necessary

Clean and well-pressed apron or uniform;

Closed and clean shoes, and that the material of their manufacture is durable,
The use of fabric sneakers or cloth flats is not allowed.
Hair must be clean, and when long, it must be tied up with
buckles and nets, according to standardization.
Nails should always be short and clean, preferably with light nail polish.
so that the subungual dirt is perceived and eliminated.
The decorations should be used in moderation, as NR32/2008 provides that the
they should not be used in everyday situations during material handling
biological, as well as in direct contact with patients. Earrings should be short and the
the use of rings is allowed.
Interferents in sample collection - direct action of collection

Evaporation of alcohol → Hemolysis

Tube sequence → Anticoagulant contamination / Result alteration
Homogenization→Coagulum
Inadequate packaging → Change of result / Stability of
samples.
 CHAPTER III: BLOOD COLLECTION

Types of blood samples

Soro

It is the liquid part of the blood obtained when collected in a TUBE WITHOUT ADDITIVE +

CENTRIFUGATION.

Plasma

It is the liquid part of the blood obtained when collected in a TUBE WITH

ANTICOAGULANT + CENTRIFUGATION.

Total Blood

In this state, the blood remains close to normal characteristics.

obtained when collected in a TUBE WITH ANTICOAGULANT, does not undergo the process of
centrifugation.
Materials used for blood collection

Types of Tubes

The collection tubes vary in size and may or may not contain additives. The additives
they either coagulate or preserve some blood component to be analyzed,
when the correct blood/additive proportion is maintained. This is why it is vital that
follow the nominal volume indicated on each collection tube, as well as the order.
for their collection.

Separator gel

Allows for complete separation between the serum and the clot,
allowing the sample stability to be prolonged;
Reduction of interferents in storage and transport;
Higher yield of the serum.

Plastic tube without separator gel

It contains silica in its composition, a natural coagulation activator;

Used for serum extraction.

Buffered Sodium Citrate 3.2%

Used for obtaining plasma;

It is used to inhibit the formation of the coagulation cascade.
EDTA K3

Used for the collection of whole blood and plasma (if centrifuged);
Inhibits coagulation by chelating calcium.

Sodium Fluoride + EDTAK2

Used for obtaining plasma (if centrifuged);

Initiate glycolysis.

Sodium Heparin or Lytic

Used for plasma extraction;

Inhibit the coagulation process;
Clear and cell-free plasma;

EDTAK2 + Separator Gel

Decrease in changes during storage and transport;

Used for obtaining plasma;
High performance for plasma production.

EDTAK2 with or without metal-free additive

Book of metals and additives;

Used for obtaining serum.

ACD (Citric acid, sodium citrate, and dextrose)

Preserves blood cells for up to 21 days (1° to 6° C).

Buffered Sodium Citrate 3.8%

Used for ESR (Erythrocyte Sedimentation Rate).

Syringe prepared with Lytic Heparin with balanced calcium

Used for blood gas dosing and other intensive care tests.

Product Portfolio - Nominal Volume (Adults)

Product Portfolio - Nominal volume (Pediatric and Neonatal)

SARSTEDT ® LEGEND

1. EDTA K Tube - Volume 500 µl. PURPLE CAP

2. Gel Infusion Tube - Volume 500 µl YELLOW CAP
3. Lytic Heparin Tube - Volume 500 µl GREEN CAP
 LEGENDA GBO ®

1. Sodium Fluoride Tubes – Volume 250 500 µl. GRAY CAP

2. Gel Tube - Volume 500 µl YELLOW CAP
3. EDTA K Tubes - Volume 500 µl. PURPLE CAP
4. Sodium Citrate Tube - Volume 500 µl or 1000 µl BLUE CAP

Minimum volume required for blood collection

We remind you that the minimum volume to be collected depends on the number of tests.
requested in that sample

BD TUBES ®:

Serum Separator Gel Tube 3.5 ml (Yellow Stopper): 1.5 ml

Gel Separator Serum Tube 5.0 ml (Yellow Cap): 2.5 ml
EDTA tube 4.0 ml (Purple Cap): 2.0 ml
EDTA tube 2.0 ml (Purple cap): 1.0 ml
PPT Tube 5.0 ml (White Stopper): 3.0 ml
Heparin Tube 5.0 ml (Green Stopper): 3.0 ml
Metal Tube with Coagulation Activator without Gel 6.0 ml (Navy Blue Stopper): 4.0 ml
Metal Tubes with EDTA 6.0 ml (Navy Blue Cap): 4.0 ml
Citrate Tube 2.7 ml (Blue Cap): 2.7 ml (total volume of the tube)
Citrate Tube 1.8 ml (Blue Cap): 1.8 ml (total volume of the tube)
Fluorescent Tube 2.0 (Gray Stopper): 2.0 ml (total volume of the tube)
ACD tube 8.5 ml (Light Yellow Cap): 6.0 ml

GBO TUBES ®:

Separator Gel Serum Tube 5.0 ml (Red Stopper with Yellow Ring): 2.5 ml
Gel Separator Tube 4.0 ml (Red Cap with Yellow Ring): 2.0 ml
Citrate Tube 3.5 ml (Blue Cap) with white ring): 3.5 ml (total volume of the tube)
Citrate Tube 2.0 ml (Blue Cap) with white ring): 2.0 ml (total volume of the tube)
EDTA tube 4.0 ml (Purple cap):: 2.0 ml
EDTA tube 2.0 ml (Purple cap with white ring): 1.0 ml
Heparin Tube 4.0 ml (Green Cap): 2.0 ml
Fluorescent Tube 4.0 (Gray Cap): 4.0 ml (total volume of the tube)
Fluoride Tube 2.0 (Gray Stopper with White Ring): 2.0 ml (total volume of the tube)
Tube with Coagulation Activator without Gel 4.0 ml (Red Stopper): 2.0 ml
Metal tube with Heparin 6.0 ml (Navy Blue Cap): 3.0 ml
Sodium Citrate Tube for VHS 1.6 ml (Black Cap): 1.6 ml (total volume of the tube)
 Sequence for Collecting the Tubes
According to CLSI Clinical and Laboratory Standards Institute (2009):
Systems for Collection

Closed System

Known as a vacuum system, it uses an adapter for a needle and

vacuum needle.

This system should be prioritized by:

Ease of handling.
Exact collection of blood volume due to the existence of vacuum
calibrated.
Patient comfort due to the possibility of collecting multiple tubes
with a single puncture.
Professional safety, since vacuum collection is a closed system.

Open System

It can be carried out using syringes in conjunction with:

needle, scalp or directly from catheter.

This type of collection should be carried out only in situations

exceptional such as:

Patients with very difficult venous access.

Client with hair fragility.
Very small children.

Open System through Catheters

The laboratory does not recommend puncturing through jelco, angiocath or

insight due to the risk of hemolysis, if it is indispensable it should -
a specific adapter should be used for this purpose.
Sources for obtaining the sample

Blood samples can be obtained from various locations in our body, among
these places are worth mentioning:

Venosa
Arterial
Capillary;
Catheter

Venous Puncture

Definition

It is an invasive method, where the venous network is accessed for the infusion of drugs and/or

solutions, as well as blood collection for laboratory tests.

Techniques for vein visualization

Ask the client to lower their arm and make opening and closing hand movements.
Gently massage the patient's arm (from the wrist to the elbow).
Vein fixation with fingers in cases of flaccidity.
Local application of heat with the aid of a hot water bag.
Observation of caliber veins.
Palpation: performed with the index finger of the collector. Do not use the thumb.
due to the low sensitivity of pulse perception. This procedure helps in
distinction between veins and arteries by the presence of pulse, due to greater elasticity
and the greater thickness of the walls of the arterial vessels.

Transillumination: a procedure in which the collector uses one or two sources

primary lights: the first, of high intensity; the second uses LED. The
cutaneous transilluminator equipment is of great assistance in locating veins,
through beams of light emitted within the patient's subcutaneous tissue. The
the user must fix the tourniquet in the usual way, sliding the transilluminator through the
skin, always adhering to the surface to prevent light scattering. The veins will be
views like dark lines. Once the best site for puncture is defined, the
transilluminator is fixed in the chosen area, taking care not to interfere with the
blood flow. There is needle insertion, completing the procedure as of
costume. The transilluminator is particularly useful in: neonates, patients
pediatric patients, elderly patients, obese patients, patients with hypotension, whose
The location of veins is difficult.

Application of the tourniquet

Position it about 8 cm or 4 fingers above the puncture site;

Should not be used for more than 1 minute;
Do not squeeze tightly, the pulse should remain palpable;
If the customer is allergic to latex, do not use this material for
strangulation.

Palpation;
Milking;
Application of heat.
Note: Do not pat the area to be punctured, especially in the elderly, as
if there are atheromas, they may dislocate bringing serious consequences.

Choice locations for venous puncture

Various locations can be chosen to perform venous puncture.

Prefer to perform the puncture in the median vein of the arm, due to it being a
vein with good visualization and good venous flow.
The other veins of the arm (Basilic and Cephalic) as well as those in the hand regions or
of the feet, can be punctured, but special care must be taken during the
collect because they are more susceptible to the formation of hematoma (mainly
the Basilica - Internal lateral vein of the arm.
Venous System

Venipuncture sites
 Places to avoid for venipuncture

Places with scars or burns;

Areas with intravenous hydration therapy; In cases where this is not
possível, coletarSEMPRE ABAIXO da infusão, o mais longe possível, e avisar ao
collection conditions laboratory.
Member close to mastectomy, catheterization, or any surgical procedure;
Previously thrombosed veins (swollen).

Venous blood collection procedure in a closed system

Inform the client about the procedure to be carried out

Position the client's arm on the support (angled down from the height of)
shadow
Sanitize hands.
Put on the procedure gloves.
Choke the client's arm/forearm
Select the vein to be punctured, release the tourniquet.
Prefer to perform the puncture in the median vein of the arm. The other veins
of the arm (Basilic and Cephalic) as well as those of the regions of the hands or feet, can be
punctured, but special care must be taken during collection because the
they are more susceptible to the formation of hematoma (mainly the Basilica -
Inner lateral vein of the arm.
Prepare the appropriate material for the puncture: multi-needle 21 or 22G.
depending on the caliber of the vein, holder and tubes for collection.

Open the needle in front of the client informing that it is disposable.

Connect it to the holder.
Strangle again the chosen member.
Perform antisepsis of the puncture site with 70% alcohol.
Let the moisture of the alcohol dry.
Remove the protection from the multiple needle

Puncture the vein.

Insert the needle into the skin with the bevel facing up at a 30º angle.
with the help of the other hand, stretch the skin below the site of the puncture

Hold the holder with one hand, preventing the needle from moving.
 Insert the first tube for vacuum collection and release the tourniquet, if possible,
moment when the blood enters the tube (follow the standardized order in item
Sequence for collecting vacuum tubes.
Homogenize the tubes about 5 to 8 times, noting that it is necessary to collect the
volume of blood specified in each container, due to proportion
anticoagulant/blood.
Remove the needle from the vein

Compress with cotton the puncture site for at least 1 to 2 minutes.

Request the customer's assistance, when possible, in the first seconds of
compression, to carry out the disposal of the needle in the perforating material collector

cutting.
Apply the adhesive bandage.
When possible, ask the client to maintain compression for 3 to 5 minutes
avoiding bending the forearm during this period (if the collection was done in this
If the puncture is in the hand, ask the client to keep their hand raised.
discreetly upwards for 5 minutes.
Recommend to the client not to carry weight or a bag with the member whose vein
was punctured for at least 1 hour.
Forward the material for collection confirmation.

Open system blood collection procedure

Inform the client about the procedure to be performed.

Position the customer's arm on the support (tilted downwards from the height of
shadow)
Wash your hands.
Put on the procedure gloves.
To garrot the client's arm/forearm.
Select the vein to be punctured, release the tourniquet.
Give preference to performing the puncture in the median vein of the arm. The other veins
of the arm (Basilic and Cephalic) as well as those of the regions of the hands or feet, can be
punctured, but special care should be taken during collection as the
they are more susceptible to hematoma formation (mainly the Basilica -
Inner lateral vein of the arm.
Prepare the appropriate material for the puncture according to the gauge of the vessel:

hypodermic needle, scalp, and syringe.

Open the materials in front of the client, informing them that they are disposable.
 Strangle again the chosen member.
Perform antisepsis of the puncture site with 70% alcohol.
Let the alcohol moisture dry.
Connect the needle or the scalp to the syringe.

Puncture the vein.

Insert the needle into the skin with the bevel facing up, at a 30º angle. With the
help from the other hand, stretch the skin below the puncture site
Release, if possible, the tourniquet at the moment the blood enters the tube
Slowly aspirate the necessary blood volume according to the amount of
blood requested for each tube.
Remove the needle
Compress with cotton at the puncture site for at least 1 to 2 minutes.
Request the client's assistance, when possible, in the first seconds of
compression, to carry out the disposal of the needle in the sharp material collector
cutting.
Open the tube cap and slowly transfer the blood, allowing it to
run along the wall of the tube in order to avoid hemolysis.
Carry out the same procedure until all tubes are collected, (Follow the
standardized order in the item Sequence for collecting the tubes with syringe.
Homogenize the tubes about 5 to 8 times, noting that it is necessary to transfer.
the specified blood volume in each bottle, due to the proportion
anticoagulant/blood.
Put on the adhesive bandage.
When possible, ask the client to maintain compression for 3 to 5 minutes.
avoiding bending the forearm during this period (if the collection was done in this
local). If the puncture is in the hand, ask the client to keep their hand raised.
discreetly upward for 5 minutes.
Advise the client not to carry weight or a bag with the limb whose vein was affected.
punctured for at least 1 hour.

Important
In open system collections, ensure that no bubbles are formed.
Do not pull the syringe plunger with too much force, nor perform the movement of
go-and-come, as the pressure may cause hemolysis.
If the medical request also includes the request for venous gasometry or
arterial, connect to the blood gas syringe on the scalp, perform the collection of this test, change

the syringe and collect the other tests.

 Hypotheses for failure:

It is possible that the needle's bevel did not fully penetrate the vein.
The puncture should be continued until the needle penetrates properly.
The needle was inserted deeply and pierced the vein. Withdraw with the
needle until the blood flows.
The needle penetrated beside the vein. Palpate the vein with the left hand and adjust.
the trajectory of the needle.

In case of collapse by the vacuum system, remove the tube and reattach it.
again, in order to restore blood flow.
In small caliber veins, it is recommended to use a fine scalp (e.g., common 25G or the
vacuum)

Arterial Puncture

Definition

It is an invasive method that uses the arterial network where a puncture is performed.
to measure the concentrations of oxygen, ventilation, and the acid-base state of the patient
critical, perform gas measurements or collect specific tests.

Choice of puncture site

Various arteries can be chosen to perform arterial puncture; however, the artery
The radial is the only artery that can be punctured by the laboratory collectors.
 Requirements for the puncture

The arterial puncture sites must meet the following requirements:

Available collateral blood flow

(Allen test);
Superficial or easily accessible location;
Tissue integrity;

Modified Allen Test

Apply pressure on the radial and ulnar pulse with your fingers to block the flow.
arterial
Ask the client to clench their hand tightly (or ask for assistance from another person to
the procedure in case of an unconscious patient) to force the blood originating from the
hand.
When the client's hand turns pale, the pressure on the ulnar artery should be
released;
The palm region and fingers are observed, in case the hand turns red inside
seconds, this indicates that total perfusion through the ulnar artery is present
confirming that it is safe to puncture the radial artery.
 Modified Allen test negative (without ulnar artery perfusion) is indicative
due to collateral circulation compromise, in this case, the radial artery will not be able to
to be accessed;

Contraindications

Change in hemostasis;
•Percutaneous puncture of any artery deeper than the radial artery (which
theoretically increases the chances of complications;

Local Compression

Local compression for 5 minutes is essential to prevent bruises and/or

ischemia should be discarded. Therefore, the fixation with microporous tape must follow
the institutional protocol.
 Potential complications of the puncture

Thrombosis of the punctured artery;

Embolism;
Infection;
Local pain

Arterial blood gas analysis

The radial artery is the only artery that should be punctured by the collectors of the
units, and the brachial and dorsal arteries of the foot should be punctured by
nurse or doctor of the unit and/or hospital.

Radial Artery

Indications for gasometry collection

The analysis of gases in arterial blood is fundamental in the treatment of patients.
critics, as it provides a quick and reliable diagnosis of insufficiency
respiratory, as well as its monitoring in relation to oxygenation, state
acid-base and alveolar ventilation.

Contraindications
Hemostasis alterations.
 Absence of central pulse.

Materials
Heparinized syringe (preferably balanced lytic heparin);
Scalp or Needle
Alcohol swab
Sterile gaze
Microporous tape

Procedure for collecting arterial blood gas

Guide the client regarding the procedure to be carried out.

Hygienize your hands.
Open the envelope of the blood gas syringe and remove the orange cap.
luer cone (syringe tip). Do not disregard this cap.
Fit the needle or common scalp into the necessary gauge (recommended in
adults: needle 25x8 / scalp 21G and recommended for children: needle 25x7 / scalp
23G) no luer cone (tip) of the syringe.
Hygienize your hands.
Put on the procedure gloves.
Feel the pulse of the artery with the index and middle fingers.
Perform antisepsis of the site to be punctured with 70% alcohol.
Puncture the artery.
Collect 2.0 ml of arterial blood.
In the case of children, a minimum of 0.5 ml can be collected.
 Remove the existing air bubbles by carefully pushing the plunger.
syringe upwards with the protection of a gauze or cotton.
Place the orange cap to seal the tube, thus preventing the entry of air.
Homoegenize the syringe 3 to 4 times. Attention: Do not homogenize before the
removal of air bubbles.

Apply intense compression to the punctured site for at least 3 minutes.

Perform a compressive dressing with adhesive tape or according to the standardization.
go to the hospital.

Identify the syringe with the barcode label.

Place and transport the material collected on the ice.
Forward the material for analysis immediately after collection.

In hospital units with a pneumatic system, it is not necessary to put the

material in the ice, due to it being quickly forwarded to the technical department.

Most common pre-analytical errors

Hemolysis;
Air bubbles;
Mix of arterial and venous blood;
Storage at room temperature for an extended period of time;
Inadequate mixing of the sample before analysis;
Failure to remove clots from the syringe tip.

Stabilization of respiratory conditions

In order to obtain a real picture of the patient's respiratory conditions, they must be
in a state of stable ventilation:

It must remain at rest for 5 minutes;

ventilation parameters unchanged for 20 min.

The pain and the state of panic from the arterial puncture can influence the stabilization of the

respiratory state and therefore should be minimized.

Mixture of arterial and venous blood

When puncturing an artery, one must be careful not to obtain,
accidentally, arterial blood mixed with venous blood. This can
it can happen, for example, if a vein is reached before the artery.

Even a small amount of venous blood can significantly change.

the results, this is specifically true for parapO2 esO2, but others
parameters may also be affected.

In the arteries, there will normally be enough pressure to fill a syringe, if

this does not happen, it may be because a vein was hit. In this case a new
sample must be taken.

Air bubble contamination

The air bubble must be expelled as soon as possible after collection with the aid of
cotton, to prevent accidents, and avoid contamination of O2.

Storage
Due to the volatile nature of gases and metabolism, the storage time
it must be at least - the ambient temperature, less than 10 minutes.

If the sample is stored for more than 10 minutes, it must be cooled (0-4
°C), to decrease metabolism.

Catheter collection

All collection procedures using a catheter, regardless of type, must be

performed by the hospital nursing team, or by the qualified nurse of
laboratory, following the standardization (internal protocol) of it

Protocol for catheter collection

According to the CLSI (Clinical and Laboratory Standards Institute) protocol, it should be
twice the volume of the catheter extension was disregarded, for this purpose was carried out

an evaluation of the catheters usually used for blood collection and we have.
The Brazilian Society of Clinical Pathology (SBPC) does not recommend that the samples be...

coagulation tests should be collected in previously heparinized catheters.

Double Lumen Catheters, Hickman, Permcath, Port-a-cath and PICC

Pediatric catheters = 3 ml of blood/fluids will be discarded before collection

blood for the laboratory;
Adult catheters = 5 ml of blood/fluids will be discarded before collection
blood for the laboratory;

Except: Porth catheter Abdominal

Adult Catheters = 6 ml of blood/fluids will be discarded before collection

blood for the laboratory;
Pediatric catheters = 4 ml of blood/fluids will be discarded before collection
blood for the laboratory;

Collection Procedures – Peripheral Venous Catheter

 Sanitize hands;
Put on a mask;
Put on the procedure gloves;
Perform asepsis with a 70% alcohol swab on the device's extension and connection.
venous
Connect the syringe to the catheter connection, aspirate the prescribed volume by
Laboratory (based on CLSI/2008) and disregard;
With another syringe, connect to the catheter and aspirate the necessary amount of.
blood for the completion of tests;
Disconnect the syringe and transfer the collected blood to the tube;
Dispose of the used syringe;
Prepare a syringe containing 20ml of 0.9% saline solution, connect and inject.
in peripheral venous access.

If peripheral venous access is maintained, heparinization should be performed.

from this access, according to institutional protocol.

Collection Procedures – Central Venous Catheter

Sanitize hands;
Put on a mask;
Put on the procedure gloves;
Perform disinfection with a 70% alcohol swab on the device's extension and connection.

central venous
Connect a syringe to the catheter connection and open the clamp;
Aspire the volume recommended by the Laboratory (based on CLSI/2008) and
despise;
Connect another syringe, open the clamp, and aspirate the necessary amount of.
blood for the completion of the tests;
 Close the clamp, disconnect the syringe, and transfer the collected blood to the tube;
Dispose of the used syringe;
Connect a syringe containing 20 ml of 0.9% Saline, open the clamp and
inject into the central venous catheter;
Close the clamp, remove the syringe connected to the catheter.

Collection Procedures – Central Arterial Catheter

Sanitize hands;
Put on a mask;
Put on the procedure gloves;
Perform asepsis with a 70% alcohol swab on the extension and connection of the device.
catheter with the PAM equipment;
Connect a syringe to the catheter connection, aspirate the prescribed volume by
Laboratory (based on CLSI/2008) and disregard;
With another syringe, aspirate the necessary amount of blood for the procedure.
of the exams;
Disconnect the syringe and transfer the collected blood to the tubes;
Dispose of the used disposable syringe;
Reinstall the PAM equipment.

Hair Collection

Definition

The capillary collection is done when a small blood sample needs to be taken from
easy, fast, and safe method, primarily used in neonates.

Procedure

Choose the appropriate location as shown in the figure below for the puncture.

Warm up the area to be punctured with a heating pad or glove.

procedure with heated water.
Perform antisepsis with alcohol and dry with gauze.
Perform the puncture with the standardized (automatic) lancet.
Remove the first drop of blood with a gauze, as it contains excess fluid.
tissular.
Leave the punctured area always below the heart and press gently.
the adjacent tissue in order to increase blood flow.
 Each drop of blood that forms at the site must be touched with the collection tube,
the drop flows into the tube by capillarity.
Do not 'scrape' the drop of blood with the edge of the tube, as this promotes coagulation.

Do not press hard, 'milking' the area, as this causes hemolysis and an increase in
amount of tissue fluid in the collected sample, producing erroneous results.
First collect the tube with anticoagulant (purple lid), then the dry tube or
with separating gel (yellow cap).
The tube with anticoagulant should be inverted about 8 times performing the
movement type 'lowering the thermometer's temperature' to remove the excess
of blood from the edges and lid.
After collecting, elevate the punctured area and press with gauze until it stops bleeding.

bleeding.
The use of the dressing at the site is up to the employee and the client.
punctuated.

Puncture sites

Lateral face of the plantar surface of the heel;

Palmar surface of the distal phalanx of the fingers;
In children under 1 year old, the heel is usually used;
In older children and adults, the most commonly used site is the phalanx (finger) of the hand;
Do not puncture the little finger and central area of the sole of the foot;

In small children, the fingers of the hands should not be punctured, as the distance
the distance from the skin to the bony surface of the distal phalanx is very small.

Examples of poorly collected material

Procedures in seated patient
Ask the patient to sit comfortably in a chair.
blood collection. It is recommended that the chair have armrests and prevent
falls, in case the patient loses consciousness. Armless chairs do not
they do not provide adequate support for the arm, nor do they protect patients in cases of

fainting.
It is recommended that, when resting in the chair, the position of the patient's arm be
slightly inclined downwards and extended, forming a straight line from the shoulder
for the wrist. The arm should be firmly supported by the rest and the elbow should not
it must be bent. A slight curve can be important to prevent hyperextension of the
arm.

Procedure on a patient in bed

Ask the patient to get into a comfortable position.
If you are in a supine position and additional support is needed, place one.
pillow under the arm where the sample will be collected.
Position the patient's arm slightly angled downwards and extended, forming
a straight line from the shoulder to the wrist.

If in a semi-sitting position, the positioning of the arm for collection becomes

is relatively easier.
Evaluation of the work done

For a plasma, whole blood, or serum sample to be of 'good quality', or

be, produce the best results, it should not contain:

Hemolysis

Abnormal erythrocyte lis with substance release, mainly

Hemoglobin. It is recognized by the reddish appearance of serum or plasma.
after centrifugation or sedimentation.

Hemolysis actively interferes in some tests, causing an increase in activity.

plasma levels of enzymes, such as aldolase, aspartate aminotransferase, phosphatase
alkaline, lactate dehydrogenase dosages of K, Mg and P.

Good practices for the prevention of hemolysis

Pre-collection
Let the alcohol dry before starting the puncture.
Avoid using needles of smaller gauge. Use this type of material only when
if the patient's vein is too thin or in special cases.
Avoid collecting blood from an area with a bruise.
In vacuum collections, puncture the patient's vein with the bevel facing up.
Perfure the vein with the needle at an oblique insertion angle of 30 degrees or
less. This way, it prevents the blood from hitting the wall of the tube forcefully,
hemolysing the sample, and it also prevents the reflux of blood from the tube to the
patient's vein.
Tubes with insufficient or excessive blood volume alter the proportion.
blood correction/additive, leading to hemolysis and incorrect results.
 In collections with syringe and needle, check if the needle is well adapted to the
syringe, to prevent the formation of foam.
Do not pull the syringe plunger too hard.
Still in collections with a syringe, discard the needle and slide the blood.
or carefully along the tube wall, making sure there is no contamination
from the end of the syringe with anticoagulant or with clot activator
content in the tube.

Do not perform the procedure of piercing the needle into the rubber cap of
tube for transferring blood from the syringe to the tube, as it may create a
positive pressure, what it causes, in addition to hemolysis, the displacement of the cork of

tube, leading to the breaking of equipment.

Pre-collection
• Smoothly homogenize the sample by inversion 5 to 10 times, according to
the manufacturer's instructions; do not shake the tube.
Do not let the blood come into direct contact with ice when the analyte is to be dosed.
need this conservation.
Pack and transport the material according to the guidelines of the Surveillance
Local sanitation, from the manufacturer's instructions for pipe use and from the manufacturer of the

diagnostic set to be analyzed.

Preferably use a primary tube; avoid transferring from one tube to another.
Outro.
Do not leave the blood stored in the refrigerator for too long before making the
exams. Check the manufacturer's recommendations for the test.
Do not centrifuge the blood sample in the tube to obtain serum before
end of clot retraction, as the formation of the clot is not yet
complete, or what can lead to cell rupture.
Do not use the centrifuge brake to interrupt the centrifugation.
tubes. This sudden interruption can cause hemolysis.

Clot

Upon detecting the presence of clots in the sample that is whole blood or
we must plasma:
 Separate the sample and request that a nurse or specialist analyze it.
sample, and confirming the presence of clots this sample should be discarded and
a new collection requested.
We must avoid the formation of fibrin so that the sample is faithful to reality.
client clinic.

Special collections

Check if the Regional conducts these tests and their references.

Bleeding Time (BT)

Procedure:

Massage the earlobe for 30 seconds.

Perform the antisepsis of the earlobe with 70% alcohol. Wait for it to dry well.
local.
Puncture the earlobe with the lancet in the lowest region.
Immediately activate the stopwatch or use a clock.
Dry the blood every 30 seconds with filter paper, touching it to the drop.
gently (do not rub).
Continue until the bleeding stops.

Coagulation Time (CT)

Procedure:

Leave the stopwatch 'hanging' around your neck.

Perform the venipuncture procedure.
The tube for performing the TC should be the last tube to be collected.
Collect approximately 2.5 ml of blood (half of the tube).
Start the timer when the first drop of blood enters the
tube.
Do not homogenize the tube.
Place the tube in a water bath at 37ºC plus or minus 0.5º.
Perform the first check in the 3rd minute or 4th minute for the PR region.
After 3 or 4 minutes, observe the presence of the clot every 30 seconds.
inverting the tube gently.
 Coagulogram

Check the test profile according to the Regional.

Procedure:
1st Bleeding Time
2° Perform the venipuncture, first collecting a dry tube (separator gel - stopper)
yellow or without additive - red cork). Release the tourniquet.
3° Collect the Citrate Tube (Blue Cap).
4° Collect the Sodium Citrate Tube 3.8% (Black Cap) when requested
of VHS.
5° Collect the EDTA Tube (Purple Cap).
6° Collect the Tube Without Additive (Red Stopper). Perform the Coagulation Time.

Thermoblock and stopwatch.

Care

Use the pediatric EDTA and Citrate tubes for collections with venous access.
difficult.
Avoid prolonged choking.
Homogenize the tube (5 to 8 times) immediately and properly after the
collection.
Do not pull the syringe plunger forcefully during the collection procedure.
Do not inject the blood into the tube with the help of the scalpel.

Do not collect from the same member that is receiving intravenous infusion.

Rope Test
 Check if the client has any type of injury on the forearm and back of the hand.
hand that may be confused with petechiae (e.g. freckles).
Check the client's blood pressure.
Calculate the average pressure as follows:

Average pressure = maximum pressure + minimum pressure

Exemplo: 100 + 70 = 85 mmHg

. Keep the sphygmomanometer on the client's arm at the average pressure for 5
minutes, always checking the pulse that must be palpable.
. Advise the customer that numbness, tingling, and cyanosis may occur.
member.
. After 5 minutes, remove the device and check immediately and again
after 5 minutes, if there was the appearance of petechiae (reddish spots on the
peel.
. Check for the presence of petechiae in an area of approximately 5 cm X 5.
cm, just below the elbow crease.
. The result must be noted in the order of execution and then it must be typed.
in the Computerized System.

Note: Clients who have undergone mastectomy, catheterization, extraction of a vein or

artery for bypass surgery and no option in the other arm, the Lasso Test not
it should be carried out; we need to contact the client's doctor.

When the client has only one arm, first carry out the Test of
Tie, wait 15 minutes and then perform the puncture.

Hypertensive patients whose average pressure is greater than 100 mm Hg, place the
device only at 100 mmHg.
 CHAPTER IV: MICROBIOLOGY

Blood culture

Material
For blood culture collection, culture bottles/media with aspiration are used.
vacuum.

Check the volume to be collected directly in the vial, remembering that this is a
essential requirement for the execution of the tests.

1 2 3 4

Blood culture Bact/Alert®.

Bact Alert Aerobic/ Yeasts

2. Bact Alert Anaerobic
3. Bact Alert Pediatric
4. Basct Alert Myco/Fungi
 1 2 3 4

BD® blood culture.

1. Aerobic BD / Yeasts
2. Anaerobic BD
3. Pediatric BD
4. BD Myco/Fungi

Procedure

Wash your hands.

Disinfect the lid of the culture medium with 70% alcohol, leaving the cotton in there until
the moment of use.
Choose the appropriate vein.
Put on the procedure gloves.
Perform the antisepsis of the puncture site with sterile gauze and Alcoholic Chlorhexidine
0.5%, in circular movements. This antisepsis should be performed through
circular movements, starting at the point of needle insertion and expanding in
circular movements towards the edges, until reaching approximately 2cm above and 2cm
below the insertion site.
Let the area dry for approximately 30 seconds.
Strangle the client's arm.
Puncturing the vein with a needle or scalpel (remember that we can no longer
lean on the arm.
Aspire the necessary volume according to each medium.
Insert the needle (the same one that was used for puncturing) in the middle, where the
blood will be aspirated by vacuum.
Homogenize the bottle with circular movements (do not shake vigorously).
Aseptic procedure for blood culture collection.

General Care

It is mandatory to record the access that was used in every culture bottle.
for the collection, PERIPHERAL - ARTERIAL - CATHETER - PORT-A-CATH.
Collect the volume specified by the manufacturer.
Note the collection time on the jar.
Record the patient's temperature (if there is a fever spike).
When requested for more than one sample, note the sample number and collect.
in places with an interval of at least 15 minutes
Each collection should be from different veins (locations).
Blood culture collection should be performed, when possible, before starting the
treatment with antibiotics.
When the doctor does not specify collection during the fever peak, the collection can be performed.

at any time of the day.

Secretion cultures

Cultura Aeróbica e de Fungos

Material
Illustration of Aymes' Middle.

Illustration of the Aymes medium thin stem.

Procedure
Put on the procedure gloves.
Before starting the collection of the requested area, clean the place to be.
collected with sterile gauze and saline solution to remove the excess secretion.
Open the culture medium wrapper and remove the swab.
Collect the secretion from the requested site. Perform the collection from the 'bed' of the lesion.

After collecting the secretion, insert the swab into the transport tube emerging.
or inside the gel

Anaerobic culture

Material

Illustration of the medio Thioglycolate.

Procedure
 Put on the procedure gloves.
Before starting the collection of the requested area, clean the location to be
collected with sterile gauze and saline solution to remove the excess secretion.
Open the swab wrapper.
I opened the lid of the culture medium.

Collect the secretion from the requested location. Perform the collection from the 'bed' of the lesion.

After collecting the secretion, insert the swab into the transport tube, immersing it.
or inside the gel, and breaking the raste.
Close the lid.

This material can also be used for tissue culture.

Note: The appropriate material for anaerobic culture is the material collected by.
aspirative puncture, using a needle and syringe, after careful antisepsis of the skin
with alcohol and Alcoholic Chlorhexidine 0.5%.

Quantitative Culture

Tracheal secretion or bronchoalveolar lavage

Material

Illustration of the mucus collection jar.

Procedure
Procedure performed in hospital units for hospitalized patients.
Material collected through aspiration (aspiration probe) should be carried out
by physiotherapists and/or nurses.
Wear the face mask and, if necessary, the eye protection;
Sanitize hands
Put on sterile gloves;
 Perform tracheal suctioning, using aseptic technique and discard this first one.
aspirated;
Use a new probe for the aspiration of the material to be collected;
Transfer the material using aseptic technique to the sterile vial, with the help of the
aspiration.
Hygienize your hands
Send immediately to the laboratory

Catheter tip culture

Material

Illustration of the Falcon ® tube.

Procedure

Put on the procedure gloves.

Perform antisepsis of the catheter insertion site with an alcohol swab.
isopropyl 70, in circular movements.
Redo the antisepsis with sterile gauze and 0.5% Alcoholic Chlorhexidine, in
circular movements.
Perform antisepsis by making circular movements, starting from the point of
insertion with the catheter and expanding in circular movements to the edges, until
to reach approximately 2cm above and 2cm below the insertion site.
Let the area dry for approximately 30 seconds.
Remove the catheter and cut approximately 2cm off the catheter with sterile scissors.
in its distal third, place in the bottle and close the cap.
Forward the material immediately to the laboratory.
Culture of secretion collected in syringe

Illustration of the correct and incorrect procedure for collecting secretion in a syringe.

•Organic materials obtained from aspiration (cavity liquids, urine, etc.),

can be sent to the laboratory in the same syringe in which it was performed
collection, but NEVER send together with the needle as a cap, so that it does not
accidents with sharp-pointed objects occur during transportation and handling
in the technical area.

Feces culture

Material

Cary-Blair medium illustration

Procedure
Put on the procedure gloves.
Open the culture medium package and remove the swab.
Dip it in the collected feces and leave it for 2 minutes.
After collecting the stool, insert the swab into the transport tube, immersing it.
inside the gel.

Bacterioscopic

Material
Matte laminate
sterile cotton swab
Materials for bacteriological collection.

Procedure to make a smear.

Procedure

Identify the transport container for slides with the barcode label.
Manually note on the label the location/region to be collected when necessary.
Identify with a pencil the matte edge of the blade with the FAP number.

Sanitize your hands.

Put on the procedure gloves.
Collect with a sterile plastic swab or a sterile fine swab the secretion.
requested and to sow (in a circular form) in 1 sheet (for each requested region).
Place in the blade transport container.
Urine culture

Material

Urine collection vials 1 and Urine culture

Procedure in Women
To orient what will be realized o exam of culture of urine.
Show the material to be used (1 red cap urine collection kit + 1 sachet)
of wet wipes).
To orient a client what she must to go until o bathroom.
You must first wash your hands well with soap and water and dry them with a paper towel.
You should sit on the toilet, spread your legs as far apart as possible and start the
hygiene of the genital area.
Clean the external part (labia majora) using wipes.
wet, with unique movements always front to back.
Despise the handkerchiefs.

Open the urine collection kit, holding the collection cup only by the outer part
(no put a hand inside do cup).
Start urination by disregarding the first jet of urine in the bowl.
Bring the collector cup close to the urethral opening (do not touch) and collect the midstream urine.

In the same. If necessary, discard the final jet in the vase.

Transfer the collected urine to the red-capped conical tube, filling it...
or up to the mark of 12 ml.
Seal the tube well and deliver it to the indicated location.
Note: The employee who receives the material must wear gloves and must identify the
tube with the barcode label in front of the customer.

WARNING: in cases of clients with motor deficits or any other type of

abnormality that prevents the client from performing hygiene efficiently, the
the employee must perform the asepsis. We should position the client
gynecological, perform asepsis of the genital area and proceed to collect in the jar as
indicated above.

Procedure in Men

Guide the customer to the restroom.

Guide to wash hands with water and soap and dry with paper towel.

You should clean the glans with wet wipes, pulling back the foreskin and
to despise the handkerchiefs.

Open the urine collection kit, hold the collection cup only by the outer part (do not
putting your hand inside the cup.

Bring the collecting cup close to the urethral orifice (do not touch) and collect the midstream.
same. If necessary, discard the final jet in the bowl.

Transfer the collected urine to the red-capped conical tube, filling it.
up to the 12 ml mark.

Seal the tube tightly and deliver it to the indicated location.

Note: The employee receiving the material must wear gloves and must label the tube.
with the barcode label in front of the customer

Procedure in children–female sex

Guide the parents or guardians regarding the material collection.

Clean the hands.

Put on the procedure gloves.

 Ask the mother or guardian to help contain the child by holding him/her.
the legs bent and exposing the genital area. Cover with the sheet.

Perform asepsis of the genital area using wet wipes with soap.
neutral (provided by the laboratory itself), starting with the labia majora,
small lips, urethral opening, vaginal introitus and anus. Use one side of the
handkerchiefs for each region.

Always proceed with hygiene in one direction, from front to back. Perform
as many times as necessary.

Adapt the collector to the genital area.

Check if there are any residues left between the adhesive and the skin, through which it could...

to urinate.

Proceed to change the collector every hour, always repeating the asepsis, until
collect the urine; not exceeding 4 exchanges. Always observe the existence of
skin irritation in the child.

Note on the appropriate form the time the collector was placed.

After collection, transfer the urine to the red-capped conical tube.

attempting not to contaminate it.

Identify the tube with the barcode label in front of the client.

Advise the mother that, since the child's skin is sensitive, the adhesive may cause irritation.

skin and even cause chafing (depending on the number of changes of the collector).

Guide the mother on the correct way to hold the child in her arms to avoid detachment.
the collector and not lose the urine.

Note: We can consider children for collector use, individuals aged 0 to 3.

years or children above this age who do not have spontaneous urination.
All girls aged 0 to 12, the collection professional himself must
perform the prior asepsis.

If the child defecates during the urine collection, the sample must be discarded.
owing to start everythingo process of asepsis again.
In children, the use of ointments for diaper rash is common; these should be removed in
its entirety before the urine collection, observing the presence of skin irritation of the
child. If the skin is already irritated, suggest to the mother to come back another day, and not
use the ointment starting the day before the exam.

Procedure in children - male sex

Guide the parents or the guardian on the collection of the material.

Sanitize the hands.

Put on the procedure gloves.

Request the mother or guardian to assist in restraining the child.

Retract the foreskin and clean the glans and urinary meatus with the wipes.
cleaned with neutral soap (provided by the laboratory itself).

Hygiene must be strict.

Return with the foreskin in place and sanitize the penis and scrotal sac.

Place the collector and check if it is well adapted.

Change the collector every hour, always repeating the asepsis, until collecting the
urine; not exceeding 4 exchanges.

Note on the appropriate form the time the collector was placed.

After collection, transfer the urine to the red-capped conical tube, paying attention to
if to avoid contaminating it.

Identify the tube with the barcode label in front of the customer.

Do not forget that in the presence of phimosis, request the inclusion of this observation in the

FAP.

Note: We can consider children for the use of the collector, individuals aged 0 to 3.
years or children above this age who do not have spontaneous urination.
For children from 0 to 12 years old, the collection professional must carry out the
previous asepsis, in exceptional cases, mainly in older children of the sex
male, we can guide the family member to perform the asepsis and collect in the bathroom.
If the child has a bowel movement during the urine collection, the sample must be discarded.

having to start everything asepsis

o process again.
In children, it is common to use creams for diaper rash; these should be removed in
its entirety before the urine collection, observing the presence of skin irritation of the
child. If the skin is already irritated, suggest to the mother that she returns another day, and not

utilize a ointment since of a eve do exam.

Children's collector
 CHAPTER V: VARIOUS COLLECTIONS

Type I urine

Material

Urine collection bottles 1 and Urine culture

Children's collector
 Procedure

Deliver the urine collection kit to the client inside the transport jar.
Instruct the customer that upon arriving at the restroom, they should sanitize their hands.

Perform asepsis of the genital area with wet wipes.

Discard the first urine stream and collect the midstream in the collecting cup.
Transfer the urine content to the yellow capped conical tube until
reach the mark of 12 ml and seal it well.
The minimum volume required to perform the test is 10 ml.
Em casos excepcionais podemos aceitar volumes menores, desde que autorizado
by the technical sector.

Midstream urine
Follow the same guidelines above, advising the client to collect only the
first stream of urine.
The volume to be collected in the first jet is approximately 10 ml.
In some cases, depending on the exam to be collected, the container should be the pot.
sterile plastic and not the urine collection kit.

Observation
There are cases where the doctor requests the collection of first, second, and third urine.
In these cases, they should ask the client to collect each jet in a container.
correctly identifying them in order.

Collection in children - female sex

Guide the parents or guardians regarding the collection of the material.

Sanitize your hands

Put on the procedural gloves.
Request the mother or guardian to help in restraining the child by holding them.
legs bent and exposing the genital area. Cover with the sheet.
Perform asepsis of the genital area with wipes moistened with neutral soap (provided
by the laboratory itself), starting with the labia majora, labia minora, meatus
urethral, vaginal introitus, and anus. Use one side of the wipes for each area.
Always proceed with hygiene in one direction, from front to back. Perform as many
times necessary.
Adapt the collector in the genital area.
Check if there are any remnants between the adhesive and the skin where it might leak.
urine.
Proceed to change the collector every 60 minutes, always repeating the asepsis, until
collect the urine, not exceeding 4 changes. Observe for the presence of skin irritation
of the child.
Note on the printed form Child Urine Collection - Collector Control, the time that the
collector was placed and the other change times.
After collecting the urine, transfer the urine to the conical urine tube with lid.
yellow.
Identify the tube with the barcode label in front of the customer.
Advise the mother that, since the child's skin is sensitive, the adhesive may irritate the skin and
up to causing chafing (depending on the number of changes of the collector).
Guide the mother on the correct way to hold the child in her lap, so that it does not come off.

collector and not lose urine.

The minimum volume required for the examination is 10 ml.
In exceptional cases, we can accept smaller volumes, as long as authorized by the
technical sector.

Note: We can consider children for the use of the collector, individuals aged 0 to 3.
years or children above this age who do not have spontaneous urination.
All children from 0 to 12 years old, the collection professional must carry out the
prior asepsis.
If the child defecates during the urine collection, the sample must be discarded.
having to start the entire asepsis process again.

In children, the use of ointments for diaper rash is common; these should be removed in
its entirety before the urine collection, observing the presence of skin irritation of the
child. If the skin is already irritated, suggest to the mother that she return another day, and not

Use the ointment starting the day before the exam.

Collection in children – male sex

Guide the parents or the guardian about the collection of the material.

Sanitize your hands.

Put on the procedure gloves.
Request the mother or guardian to assist in restraining the child.
Retract the foreskin and perform hygiene on the glans and the urinary meatus with wipes.

cleaned with neutral soap (provided by the laboratory itself).

Hygiene must be strict.
Return the foreskin to its place and perform asepsis of the penis and scrotal sac.
Place the collector and check if it fits well.
Replace the collector every 60 minutes, always repeating the asepsis, until
collect the urine. Note on the form Urine Collection for Children - Collector Control,
hora que o coletor foi colocado e os demais horários de trocas.
After collection, tightly close the collector and place it inside the sterile container.

Identify with the barcode label in front of the customer.

Do not forget that in the presence of phimosis, request to include this observation in the FAP.

The minimum volume required for the examination is 10 ml.

In exceptional cases, we can accept smaller volumes, provided authorized by
technical sector.

Note: We can consider a child for the use of the collector, individuals aged 0 to 3.
children above this age who do not have spontaneous urination.
All children from 0 to 12 years old must be collected by the professional themselves.
previous asepsis.
If the child has a bowel movement during the urine collection, the sample should be discarded. If the skin is already...

Irritated, suggest to the mother to return another day, and do not use the ointment since.
eve of the exam.

24-hour urine

Material

24-hour urine collector.

 Procedure

The 24-hour urine should be collected in a proper container. The client should be advised.
to take the vial and the instruction/orientation for collection at the laboratory.

For each 24-hour urine test, there is a specific bottle with or without
preservative.
The computerized system and the 24-hour urine table provide the data
necessary for the choice of the vial and some necessary observations for the collection.
The collection vials are labeled with tags that contain information.
regarding the type of preservative, specific care with the solution contained in
bottle, the date it was produced and the expiration time.

Feces

Material

Stool collector with and without preservative

Procedure

There are several types of tests, for which the requested material is stool.

All exams are described (prerequisites, collection method,

material conditioning, delivery time, diet and general considerations) in
Computerized System or in the Collection Guidelines.
Note: Some stool tests require a prior diet: Coprological
functional, Fat Dosage in Stools and Occult Blood.
 THEORETICAL FRAMEWORK

I. BONINI, P. Errors in laboratory Medicine, 2001;

II. CLINICAL CHEMISTRY, 48:5 691-698, 2002;
III. CLINICAL CHEMISTRY, 43: 1348-1351, 2007;
IV. CLSI. Clinical and Laboratory Standards Institute, available at <
[Link] >;
V. GUDER, W.G., NARAYANAN, S., WISSER, H., ZAWTA, B. Samples: From the
patient to the laboratory, 1996;
VI. INMETRO. Good Practices of Clinical Laboratories BPLC. Qualitymark Publishing, 1998;
VII. PLEBANI, M. HEMOLYSED, K. Specimens: A reason for rejection or a clinical
challenge?, 2003
VIII. ROTH, E. How to Implement Quality in Clinical Laboratory - The Path of
Stones. Hinsdale Publishing, 1998;
IX. SIG: Integrated Management System, in the current version;
X. TESUTO, M. Manual of Clinical Microbiology, 9th Edition, 2007;
XI. WITTE, D. E. Mistakes, blunders, outliers, or unacceptable results: how many?
Clinical Chemistry (1997) 43: 1352 -1356;
XII. WATSON, D. Wrong Biochemistry results. BMJ, 324-422, 2002.
XIII. SBPC, Recommendations of the SBPC Venous Blood Collection, 2009
 ANNEXES

PRE-ANALYTICAL INTERFERENTS IN EMERGENCY LABORATORY TESTS

Stage of Action
Possible sources of errors
process preventive

Medical Request Failures in communicating the desired information

Errors in the synonymy of the requested exams, lack of Medical Request

detailed information such as desired time for standardized
collection, urgency need, communication failures Medical Prescription

verbal or written (applicant's letter) computerized

Registration of data Failures in the transcription of received information

of the patient and of
exams in the system
computerized
Electronic interface
Errors in the patient or exam registration data between the LIS of
requested. Results that are calculated based on laboratory and the LIS
no weight and height of patients (for example Clearance of do hospital. Request
creatinine) or that require the information of the date electronic doctor
time of the last dose of medication (drug dosage) with fields of
therapeutics). filling
mandatory.

Identification of the pre- Sexo, Idade, Raça, Ciclo Menstrual.

necessary requirements
for the execution of the
testes
Intrinsic influences such as sex, age, and race can Values of
change the concentration of some analytes in the blood and reference
urine. The physiological changes of pregnancy - increase suitable for
no plasma volume, increase in glomerular filtration sexo, idade, fases
and an increase in the plasmatic concentration of proteins of the menstrual cycle and
carriers modify the values of normality gestational state.
for several biochemical parameters.

Fasting

The intake of food or liquids can alter the Make sure that the
values obtained in the plasma dosages of some prerequisites
analytes, especially glucose, cholesterol and the necessary for the
 triglycerides. Urinary dosages should always be exams were
assessed as quantity per day and not quantity correctly
per liter, with the aim of minimizing variations defined,
resulting from water intake. oriented and
fulfilled.

Exercise

The acute changes produced by exercise occur Make sure that the
as a result of the changes in concentrations prerequisites
hormonal (increase in concentrations of epinephrine, necessary for the
norepinephrine, glucagon, cortisol, ACTH and decrease exams were
in insulin concentrations), loss of volume through correctly
sister and changes in the intra and compartments defined,
extravascular. These hormonal changes can, by oriented and
example, increase the leucocytes by up to 2500/mm3. fulfilled.
During training periods, the increase of
aerobic capacity is a result of the increase in
number of mitochondria, thus increasing the
muscle's ability to metabolize glucose, the acids
fatty acids and ketone bodies. These individuals have thus a
proportional increase of the MB fraction of
creatine phosphokinase, reaching up to 10%.

Identification of Medicines
medications in use

The list of
interferents is
big and complex.
The interference can be biological or chemical. Biological Existing suspicion
for example when the drug increases levels of investigate all the
plasma proteins, resulting, therefore in a possibilities
increase in serum levels of analytes linked to known.
these proteins. Chemical interference occurs in vitro, Identify the
in the same way as in hemolyzed samples or main
icteric. medications in
body use
clinical and disseminate the
main tests
who suffer
 changes with
these medications.

Identify the Fever causes hemoconcentration, altering cell counts, increases the
clinical conditions creatinine, cortisol, and uric acid.

The presence of intense pain or polytrauma raises serum concentrations of

some hormones like insulin, cortisol, renin, and growth hormone.

Prolonged bed rest causes hemodilution and reduces the

potassium concentrations, albumin and protein, through bone reabsorption increases the
ionized calcium.

Patients who are admitted to the OS after performing intense exercises may
present elevated lactate and CK values and hypoglycemia.

Collection of materials Collection time

necessary

Ensure that the

The influences of the moment of collection on the results of
prerequisites
exams may be due to the circadian rhythm (for
necessary for the
example, cortisol, renin and some markers
the exams were
tumors) to seasonal variations (for example, increase
correctly
25-OH colecalciferol in summer) or the variations
defined,
biological (for example, aldosterone in the cycle
oriented and
menstrual.
fulfilled.

Other therapeutic or diagnostic procedures

The contamination of samples by infusion solutions Collections must be

administered to the patient is the main cause of done on
pre-analytical interference in the laboratory. Never opposite member to
samples must be collected for the laboratory from veins from the infusion

close to medication infusion sites. If the intravenous.

samples will be collected from intravenous catheters or Catheter collections
intra-arterial, the cannula should be washed with saline. they must follow
physiological, with twice the volume of the catheter. Next, rigorously as
aspirate 5 ml of blood and discard. Only then the sample instructions on the side.
the laboratory must be collected. For tests of
coagulation must disregard a volume equal to 2
times the volume of the catheter, collect the necessary blood
for the completion of the other requested exams and only then
collect the sample for coagulation tests.
 If possible, after the
The infusion of drugs, contrasts, and ionizing radiation injection of
acts in many ways unknown in the contrasts
serum concentrations of some analytes. The contrasts intravenous
Radiological exams alter urine density. The hyper wait for
hydration for the performance of ultrasound promotes dilution less than 24 hours
urinary. before performing
other exams.

Blood transfusions

After transfusion
The greater the volume of blood transfused, the greater
wait near
the possibilities of changes: Hemolysis with
6 hours for collection
consequent increase in potassium and DHL.
of exams

Posture

In the standing position, the filtration pressure increases Follow the

reducing plasma volume by up to 12% in pre-instructions
normal individuals and therefore concentrating cells, analytical of
macromolecules and small molecules linked to prior rest to
proteins. The most evident changes occur in the collection of some
serum concentrations of Aldosterone, renin, epinephrine exams.
and norepinephrine.

Strangulation

The tourniquet is applied to assist the professional with Maintain

nursing the visualization of the vessel that will be used garrotting
for blood collection. The blockage of venous return brief (less than 1
what is caused by this action increases the pressure minute) for collection
capillary increase, causing liquids and small of blood. For
moléculas sejam transferidos para o espaço intersticial. collection of
The blood cells and macromolecules that do not gasometria, lactato
they manage to cross the capillary wall, then they remain in the the ideal is ammonia
glass with reduced volume and therefore it has its collect without
increased concentration. The main analytes that strangulation.
altered with prolonged strangulation are: CK, DHL,
GGT, TGO, TGP, bilirubin.

Collection location

The differences in serum concentrations in the blood Identify in the report

arterial / venous or capillary can be significant the arterial collections and
(>10%) for some analytes such as lactate, pO2 and capillaries.
bicarbonate. Hair collection is greatly influenced by
technique used, which may present differences
important in the counts of leukocytes and platelets and
hemoglobin and potassium dosages. When this
 the technique is applied, the location should be recorded in the report
puncture.

Blood / anticoagulant ratio

It is necessary to collect at least 2 times the amount. Respect the

necessary for analysis and never the total volume of the tube, blood proportions
because this can harm the homogenization of the material. anticoagulant
recomendadas.

Patient identification

Confirm the name

complete
It must be carefully checked that the material is
identify the
being collected from the correct patient, primarily in
patients
closed sectors where there is more than one bed per
interned with
fourth.
bracelet with code
of bars.

Homogenization of the material (sample x preservative)

Homogenize
The appropriate mixture of blood and anticoagulant is adequately the
essential to prevent the formation of clots material logo after
the collection.

Storage of Transport time and conditions

samples for
transport

When the urgent laboratory is located inside Store and

in this aspect, the hospital does not pose a problem. transport the
The emergency menu tests most influenced by samples according to
time interval between collection and processing is the respecting the
gasometries and the dosages of ammonia and lactate. A time
sample storage, however it must be carried out temperature and
in a standardized way, so that there is conditions
need to re-analyze the interference of light, preconceived.
evaporation, chemical reactions, and cellular metabolism
be minimized. In general, after the
processing, the serum and plasma samples must be
kept in the refrigerator, except for the tests of
coagulation, which is like total blood samples
they must be kept at room temperature.

Preparation of samples Sample preparation

for analysis Centrifugation

Wait 30
It should be done after the blood has clotted (about 30 minutes to the
minutes after collection), in speed/time blood coagulates and
predicted. Cases of insufficient centrifugation of so spin
samples can lead to altered results, the samples for 10
mainly in coagulation tests. 15 minutes to
2500-3000 rpm.

Identification of Lipemia
interferents
analytical

Serum and plasma samples may show Report in the report

varied degrees of turbidity due to the increase of that the dosage was
lipoprotein content. The presence of this lipemia is harmed by the
always a sign of abnormality and should be reported in presence of lipemia
report of the exam. The distribution of water in the samples is in the sample.
altered, interfering with the electrolyte results and
metabolites. The turbidity of the material interferes in all
the colorimetric dosages (mainly creatinine,
uric acid, CK, protein and glucose.

Jaundice

Report in the report

The presence of bilirubin in the sample affects the dosage that the dosage was
of various colorimetric tests, especially the harmed by the
creatinine dosages in most methodologies presence of
equipment bilirubin now
sample.

Endogenous antibodies

How cold agglutinins affect cell counts of Suspecting of

erythrocytes, leukocytes, and platelets. The EDTA antibodies cryoagglutinins,
dependents falsely reduce the counts put the tube in
platelets (pseudoplatelet deficiency). The macroenzymes water bath at 37
enzymes forming complexes with immunoglobulins for 15 minutes and
have been demonstrated for practically all of the process
relevant enzymes. The most frequent example of this again to a
process is the macro CK, promoting elevations in the sample.
dosages of CKMB. Autoantibodies against hormones
tireodianos are well known in clinical practice and
antiphospholipid antibodies can elevate the results
of the TTPA.
 Hemolysis

Avoid during the

collect the following
procedures:

It is the result of hemolysis, releasing its Strangulation

content in the serum/plasma, which then becomes prolonged
reddish. The effects of this process on the dosages
Biochemical processes involve the release of constituents. Rapid aspiration
cells in serum (raising serum concentrations of or with a lot
potassium and DHL). Optical interference due to color pressure.
serum redness interferes with all measurements
colorimetric. Vigorous infusion
of blood in the tube.

Blood infusion
no tube through the
needle

PARTICULARITIES OF COAGULATION TESTS

For most coagulation tests, a tube with anticoagulant is used.

universally or sodium citrate at 3.2%, in a ratio of 1 part anticoagulant to 9
blood components.

If a smaller volume than recommended is collected, the concentration of the anticoagulant increases.
and the TTPA values are falsely prolonged.

In patients with a hematocrit greater than 55%, the plasma volume is reduced and the ratio
anticoagulant - plasma is also affected. This factor falsely increases the values of PT and
TTPA.

The presence of hemolysis can activate coagulation factors and lipemia and jaundice of the sample.
You can change the measurements on the photo-optical equipment.

During the transport of coagulation samples, the containers must be kept at temperature
environment since the ice can activate factor VII and reduce the PT.

In difficult collections, coagulation is activated before we place the blood in the container with
anticoagulant, thereby altering the results. When in doubt, the collection should be repeated.

PARTICULARITIES OF HEMATOLOGY TESTS

For the analysis of blood cells, the recommended anticoagulant is dipotassium EDTA.
It is recommended to process the samples in automatic cell counters within 24 hours.
hours after collection. Immediately after collection, the tube should be inverted several times to promote the
adequate homogenization of the sample, thus avoiding the formation of clots. For a
Proper homogenization requires leaving about 20% of the tube empty.

EDTA has a great influence on the stability of leukocytes, especially neutrophils.

and monocytes. After a few hours, the degeneration of the neutrophils is evident, with
disappearance of bridges between lobes and cytoplasmic granulation. When it is collected
a sample with a volume lower than the recommended, the relative increase in the concentration of EDTA
produces evident alterations in the morphology of neutrophils and decreases the volume of erythrocytes
(reduces VCM and Htc).

Pseudoplateletpenia is a well-known event that occurs due to agglutination or

adherence of platelets to neutrophils (satellitism). Patients who demonstrate Satellitism
EDTA-induced platelet counts must have their tests collected in citrate and not EDTA.
PARTICULARITIES OF BIOCHEMISTRY TESTS

The biochemical interferents are specific methods and equipment. Thus, the results obtained
in an analytical system cannot always be directly compared to another. Sometimes
the interferents of a system do not affect another methodology.

The frequent interference of medications in creatinine levels is remarkable.

especially antibiotics (Cephalosporins).

If drug interference is suspected, a new sample should be collected immediately.

before the next dose of the medication looking for the lowest serum level to minimize the
interference in biochemical assays.

PARTICULARITIES OF BLOOD GAS TESTS

Heparin is the anticoagulant of choice for blood gas determinations.

the heparin-blood ratio must be maintained and the sample must be collected in anaerobiosis,
avoiding contact of the sample with the air or the presence of bubbles in the material (which can
result in false elevations of pO 2).

The metabolism of blood cells consumes O 2 and, therefore, the collected material must be
analyzed in no more than 30 minutes.

If transportation of the sample is necessary, it should be placed on ice and processed within 2
hours after collection.

Since sodium heparin has a very low pH, high concentrations of this anticoagulant
they can change the pH and pCO2 of the sample.

Moreover, samples collected with sodium heparin are obviously not suitable for
plasma Sodium and Potassium dosages.

Currently, syringes with lyophilized lytic heparin allow for greater control over the
heparin-blood ratio, but the homogenization of these samples must be rigorous for
prevent the formation of clots.

PARTICULARITIES OF THERAPEUTIC DRUG DOSAGE TESTS

Most of the assays are performed in serum or plasma.

After dose adjustments, it is necessary to wait a period of 5 half-lives of the drug to collect again.
sample for serum dosage.

For basal concentration determinations, samples must be collected immediately.

before the next dose (the minimum serum level of the drug is monitored).

For peak concentration determinations, respect the pharmacokinetics of each drug and route of administration.
administration (used to assess toxic levels of the drug).

Particularities of Immune/Hormone Tests

Most immunochemical tests are performed on serum or heparinized plasma.

One must consider the changes in hormone concentrations caused by daily variations and
by posture. Other hormones such as insulin and peptide-C remain stable if kept in
ice.