LABORATORY PROTOCOLS Code

SAMPLING AND STANDARDS Version 02

GENERALS
IN THE HANDLING OF THE SAMPLE Page 1 of 31

BIOS CAM DOCUMENT HISTORY

Version Date Nature of Change

before update
update
01 June 2009 All the reviews and updates are made
July 2011 biosecurity procedures that must be
apply at the SOMER clinical laboratory.

WE COMMIT TO THE APPLICATION OF THIS MANUAL

Date Readable Name Company

Elaborated and/or updated Reviewed Approved

Section Chief of Bacteriology: Quality Coordinator Medical Director

Name: ESILA GOMEZ FABIO RESTREPO Fabio Restrepo

Date June 2009 August 2009 August 2009
 SOMER CLINICAL LABORATORY

SAMPLING AND GENERAL STANDARDS

IN THE HANDLING OF THE SAMPLE

SCIENTIFIC DIRECTION
 TABLE OF CONTENTS

GENERALITIES
IMPORTANCE OF SAMPLING
2. RECOMMENDATIONS FOR ATTENTION
3. GAIN SKILL, CONFIDENCE, AND PROFESSIONALISM

4. BASIC INFORMATION AT THE SAMPLING SITE.

5. IDENTIFICATION OF PATIENTS OR USERS OF THE LABORATORY.

6. BIOSAFETY STANDARDS AT THE SAMPLING SITE.

7. USE OF ANTICOAGULANTS

8. HOW TO PERFORM VENIPUNCTURE

9. SAMPLE COLLECTION FOR CLINICAL CHEMISTRY

10. SAMPLE COLLECTION FOR HEMATOLOGY

11. SAMPLE COLLECTION FOR IMMUNOLOGICAL AND SEROLOGICAL TESTS

[Link] FOR MICROBIOLOGY AND MICROSCOPIC STUDIES

13. SAMPLING IN PARASITOLOGY

14. SAMPLING FOR MICROBIOLOGY (CULTURES).

15. SAMPLING FOR CERVICAL CYTOLOGY VAGINAL

16. SAMPLING FOR SPECIALIZED TESTING

17. TRANSPORT AND SHIPPING OF SAMPLES

18. REVIEW OF ONCOLOGY SAMPLES (FLOW CYOMETRIES, BIOPSIES,

KARYOTYPES, MYELOGRAMS, SPECIAL STAININGS E
IMMUNOHISTOCHEMISTRY

BIBLIOGRAPHY
The quality in health is given by the set of technical, scientific, and human characteristics,
financial and material resources that the Social Security in Health must have, under responsibility
of the people and institutions that make up the system and the proper use of the services
by the users.

The main characteristics of the quality of health care are: accessibility,

opportunity, security, and technical rationality. Quality integrates additional characteristics
such as professional suitability and competence, the availability and sufficiency of resources,
effectiveness, efficiency, integrality, continuity, humanized care, and satisfaction
"from the user with the attention received." Art. 3 Decree 2174 of November 1996.

A good result depends on a good sample

0.1 Activities, functions, and attitude in sampling:

The sample collection at the SOMER clinical laboratory will be coordinated by a

plant bacteriologist in whose shift schedule of the month the programming of it will appear
Responsibility. The hours of operation will be from 6:45 a.m. to 10:00 a.m. and will be dedicated
during this time exclusively to these functions, monitoring, supporting and coordinating everything
the sample collection carried out by laboratory and/or nursing assistants according to
the following parameters:

Attend to patients and/or users of the laboratory in a courteous and polite manner.

[Link] to all patients and individuals who request information in a cordial and courteous manner.
about the activities specific to the laboratory.

3. Know and apply the administrative procedures for patient care of the
different
 EPS, contracts and individuals.

[Link] correctly the demographic data and others of the patients attended,
include full names, age, sex, citizenship ID number,
phone and membership or EPS affiliation number.

[Link] economically to the clinic's administration regarding the received funds

for
concept of personal services and other payments.

[Link],record, take, receive, and transport the samples that will be processed in the
laboratory or
travel to the contracted sampling sites as required.

[Link] comply with the Biosecurity Standards established by the Laboratory.

8. Carry out centrifugation processes, staining of slides, mounting of coprology samples and
urinary sediments and other activities typical of the sample center.

[Link] with Waste Disposal Regulations.

[Link] complete the requested records and statistics daily.

[Link] refrigerators, shelves, furniture, and generally the areas in perfect order and cleanliness.
assigned work sections; collect dirty material and dispose of it properly.

[Link] and organize the special material of each work section in the laboratory.

13. Arrange the laboratory material that will be recycled in accordance with the pre-
sterilization, washing, rinsing, drying, packaging, and sterilization.

[Link] enough daily use material arranged for your disposal such as:
torundas
cotton-wrapped and sterilized, gauze, swabs, tongue depressors, plates, slides
etc.

[Link] for minimum quantities of materials needed for care.

patients of
according to the volume attended.

[Link] for the inventory in your charge.

[Link] with the schedules and programming shifts for Saturdays and rest days.
 [Link] with the general standards of organization and operation of the laboratory and
others
internal regulations of the Laboratory.
.

1. IMPORTANCE OF SAMPLING

• It is almost always the first and perhaps the only contact with the patient, user, or client.
• The image that is projected is the one that remains fixed in the people attended.
• The great responsibility in sampling extends from the quality of the
humanized attention such as the elimination of technical errors that influence the results
finals.
• The quality of the sample obtained guarantees a good final result for its analysis.
• The responsible work of each assistant who takes samples guarantees and justifies the
trust placed by the professional staff and the laboratory, as they manage a
countless specimens that entail an ethical, professional, and judicial responsibility
that compromise the entire work team.

[Link] FOR CARE:

When carrying out any sampling procedure, please take into account the following
recommendations
Act with as much kindness and politeness as possible.
Ensure that the patient is as comfortable as possible.
Transmit confidence, security, and professionalism during the procedure.
C o l l e c t t h e s a m p l e w i t h t h e b e s t q u a l i t y.
Obtain enough specimens according to the requirements of each analysis
to practice.
Permanently demonstrate attitudes of collaboration.
Demonstrate undeniable biosecurity actions.

3. GAIN SKILL, CONFIDENCE, AND PROFESSIONALISM:

Know and learn perfectly the procedures and techniques that will be carried out.
Gain confidence as your experience increases.
Develop skills in communication with patients.
Understand that you are not exempt, even in the best conditions, from making mistakes.
during some procedure. Making a mistake should promote a learning attitude and
does not constitute a fact of failure, the mistake made must be corrected so that ideally it does not
happen again.
Maintain a calm attitude.
 Think: "I have carried out this procedure many times and I will continue to do it well many"
times more.
Do not convey attitudes that denote haste, eagerness, or worry.
Make sure that each patient feels important to you.
Every attitude qualifies its degree as 'professional'.
Smile kindly.
•Wear conservative and clean clothes. Reflect adequate personal hygiene. Your uniform,
Lab coat and shoes must be in good condition.
If you have long hair, try to keep it tied up.
Your hands and nails must be kept in optimal hygiene conditions.
If you use gloves, make sure they are the right size.

4. BASIC INFORMATION AT THE SAMPLING SITE.

When attending a branch of the laboratory, check that the following are present
documents, as they are mandatory to be present, will be requested by the people who
they carry out the quality monitoring of each of the EPS and IPS.

Sampling manuals.
Procedures manuals.
Coloration Manual.
Sample collection instructions for patients.
Biosafety Standards.
Waste management and disposal.
Price lists.
Management of the different paperwork of the EPS.
Sample collection schedules.
Daily record of patients and the total number of tests to be carried out and by EPS.

5. IDENTIFICATION OF PATIENTS OR USERS OF THE LABORATORY.

When attending to any patient or user, the following information must be recorded
identification
Full names.
Full last names (2)
Age.
Sex.
Identity card number.
Telephone.
EPS membership number. (Card number).

NOTE: All these data must be recorded in the routine work notebooks of each
branch in the central laboratory.
[Link] guidelines at the sampling point.

Personal and general protection:

The use of an apron or laboratory coat is mandatory during the daily sampling shift.
sample.
The use of gloves is mandatory during the collection and handling of samples for collection.
A different pair of gloves should be used for each cytology.
Never attempt to repack the needles in their cap, use the guardian or, failing that
arrange them in the bucket with hypochlorite
The use of a mouth cover or mask and glasses is recommended for taking cytology.
The use of the refrigerator to store personal food that belongs to others is prohibited; this is a place
potentially pathogenic even with routine washing and disinfection.
It is completely forbidden to eat any type of food inside the examination venue.
samples, for this they must go to the cafeterias of each branch or failing that
do it outside the workplace.
Avoid contact of skin or mucous membranes with blood or other secretions from all
patient, all samples are potentially disease transmitters (HIV, Virus
Hepatitis B, etc.).
Wash your hands immediately before and after performing any procedure.
the branch. Remember that gloves are not substitutes for hand and skin washing.
due to the variability of its quality.
Always avoid bringing your hands to your face, mouth, and eyes when taking samples.
No smoking or wearing makeup in the work area.
• Any puncture, wound, and/or possible contamination with biological liquids or material
contaminated, it must be reported to the laboratory to be handled as an accident
work, in which case the laboratory will be exempt from all responsibility.
Record the temperature of the refrigerator and the time you did it in the form. Do it.
preferably upon arrival and always the first time I open the fridge.
•Perform a monthly cleaning of the refrigerator with 0.5% sodium hypochlorite and record in the
temperature sheet.

THE OCCUPATIONAL RISK OF HEALTH WORKERS IS MINIMAL IF KNOWN AND

RESPECT THE EXISTING BIOSAFETY REGULATIONS
[Link] OF ANTICOAGULANTS:

There are multiple factors involved in the blood coagulation process. The
Anticoagulants are substances that prevent the formation of clots. There are different
types of them in powder or liquids. The appropriate anticoagulant must always be selected
according to the study that is to be conducted.

7.1EDTA:
Ethylenediaminetetraacetic acid. This type of anticoagulant is mainly used when
studies are conducted where cells are counted as in hematology; also for the
hemoglobin electrophoresis.

7.2 SODIUM CITRATE:

Generally at concentrations of 3.8% and commonly used in studies of
coagulación: Tiempo de Protrombina, Tiempo de Trombina, Tiempo de Tromboplastina Parcial
Activated, Antithrombin III, Lupus Anticoagulant, Fibrinogen, Plasminogen, among others; these
samples must be separated within a time no greater than one hour and the plasma placed in a
plastic tube and kept under refrigeration. See Coagulation tests.

7.3HEPARIN:
It is used in some routine studies as well as specialized ones. Its presentation may include
sodium or lithium concentrations. In general, heparin with lithium is used for studies of
Chemistry and sodium heparin is used for lymphocyte studies (e.g. HLA).

7.4 OXALATES:
They are less commonly used anticoagulants, occasionally used in determinations of
glucose.

There are known international color codes for the different presentations of
blood sample collection tubes, which contain a specific anticoagulant:

Red Cap...........................No Anticoagulant (Dry Tube).

Violet Cap.......................With EDTA.
Blue Cap...........................With Sodium Citrate.
Green cap (or white)........With Heparin.
Gray Cover............................With Oxalates.

You can also obtain tubes that have a gel or separator filter feature.
serum, forming a barrier between the separated serum and the blood cells. It is found in
commercial form as 'Serum separator tubes' (SST).
8. HOW TO PERFORM VENIPUNCTURE

There are two types of time in exams. The first time (simple time), when you
it is about an exam that only requires a puncture. The second time (multiple time),
when the exam requires and includes different samples at different times. In
in either case, the necessary instructions should be given to the patient regarding
preparation, taking medications, how to wait to take second samples, etc.
To carry out a correct blood sampling, clearly follow the following
instructions:

Patient identification, call them for blood draw.

Full names and surnames, reception code and age.
Fasting verification, treatments.
Inform the patient about the procedure to follow, reassure them.
Identify the elements for sample collection. (tubes, plates, bottles, etc.)
Place the patient in a suitable (safe and comfortable) position.
Never perform blood puncture on a patient who is standing.
Prepare the materials: tubes, tourniquets, cotton, alcohol, syringes, needles, adapter
of venojet, etc.
Put on the gloves.
Gently make the patient close their fist to palpate the vein.
Select the appropriate vein, the most visible and palpable.
Do not choose a limb where an intravenous catheter is placed.
Clean the venipuncture area (with cotton soaked in iodinated alcohol) starting
from the central point and outwards.
Apply the tourniquet for no more than 30 seconds.
Firmly secure the vein.
•Perform the puncture at a 15-degree angle; follow the direction of the vein with the needle;
do not bury the needle.
Release the tourniquet when the blood flows into the tube.
After the sample is extracted, instruct the patient to relax the fist and not to pump with the hand.
Place a cotton ball without alcohol, applying gentle pressure on the puncture site.
Gently mix the tubes with anticoagulant to avoid hemolysis.
Observe the patient's condition. (dizziness, bleeding, etc.)
Provide instructions on delivering results.
Properly dispose of contaminated material.
Record the time of sample extraction.
To avoid contamination of the samples during the puncture, it is recommended, extract
first the tubes without additives and then the tubes with CITRATE and EDTA.
At the end of the procedure, instruct the patient to keep the limb bent.
with the cotton, for a time of more or less three minutes to avoid bleeding again.
• Remember: An excellent sample is synonymous with a truthful and reliable result.
quality.
[Link] COLLECTION FOR CLINICAL CHEMISTRY.

Clinical chemistry includes a large number of laboratory tests to evaluate different

organs and functions; but the most commonly indicated ones for assessing or monitoring cases
generalized or specific clinical signs are:

Glycemia Transaminases (GOT-GPT) Amylases.

Lipid Profile Total and Direct Bilirubin Ionogram. (Cl, Na, K.)
Uric acid Albumins Lactate Dehydrogenase.
Urea Alkaline Phosphatases Magnesium.
Creatinine Acid Phosphatases
Phosphorus. Prostatic Phosphatases

For them, blood must be drawn in a dry tube (without anticoagulant) to obtain a volume.
about 5 to 10 ml of serum for each test, if there are many tests to
to carry out, two or more tubes should be bled; when collecting the blood, let it coagulate to
then take to centrifugation.

9.1 GENERAL INSTRUCTIONS:

Minimum fasting of eight (8) hours. The fasting should not exceed twelve hours.
Avoid physical exercise, especially hours before the sample collection.
Patient rest for 10 to 15 minutes before the blood draw.
Do not consume alcohol the day before
No smoking before the sangria.
Ask the patient what drugs they are taking, note them down.
Do not apply the tourniquet for more than 30 seconds.
If it is an emergency, note that fasting is not required.

9.2 SPECIAL CONDITIONS:

9.2.1 Glucose Tolerance Curve:

A strict fast of eight (8) hours is required before the test.

The patient must remain at rest during the test and should refrain from drinking.
liquids, smoking, eating and exercising.
Draw blood from the fasting patient.
•Perform a glucosuria or a glycemia using a GLUCOMETER or with DESTROSTIX strips,
to verify that the patient's basal glycemia is less than 140 mg/dl or that they have a
negative glucosuria.
 In case the patient presents hyperglycemia, refer immediately to the doctor with
a note of the observed value.
Administer the required glucose load (75 or 100 grams, 50 if it's for screening), in a
a lapse no greater than ten minutes.
Prepare the glucose load of 100 and 75 grams in 300 ml of water, and the glucose load.
50 grams in 200 ml of water.
The time between each puncture must be exact: 1 hour -- 2 hours -- 3 hours -- 4 hours.
In the post-prandial blood glucose after breakfast, the first sample for blood glucose is taken.
fast and the patient is instructed to have a breakfast rich in carbohydrates (honey, sugar,
jam, bread, toast, arepa, etc.) and return to the laboratory exactly at two.
hours to take the postprandial sample.
In the postprandial blood sugar after loading, the first sample for blood sugar is taken at
fasting and a 75-gram glucose load is given to the patient and the post-sample is taken
pandrial exactly at two o'clock.
The O'sullivan test or screening for pregnant women is taken with the first sample in fasting.
A load of 50 grams of glucose is given, then a second sample is taken after one hour.
accurate for quantifying post-load glucose.

[Link] GLUCOMETER MANAGEMENT

Perform asepsis on the patient's finger that will be punctured with cotton and alcohol.
Take the aluminum envelopes out of the box and separate one.
To open the aluminum pouch. Carefully pull on the ends. Fold back.
the ends of the test strip that are inserted into the meter.
Insert the end of the test strip between the two parts of the open aluminum envelope, and
insert the strip completely into the meter.
A beep will sound and the image 88.8 will be displayed on the screen, followed by a number.
function (F#) and the results of the previous test. The function number and the results
The previous test starts to blink alternately. Remove the metallic paper.
from the test strip. Proceed immediately (within a period of three minutes) to the
next step:
Prick your finger with a lancet or an automatic lancing device and press.
until obtaining a small drop of blood.
Bring the external end of the test strip closer to absorb the drop of blood until
let the meter beep. The test strip automatically draws in blood. The timer
the countdown begins starting from 29 seconds.
Once the 29 seconds have passed, the result of the blood glucose test appears in the
screen. The test results remain on the screen.
If you want to save the test results, remove the test strip from the meter.
using a handkerchief. The meter turns off automatically and the results are
they are stored in memory.
If you do not wish to save the test results in memory, remove the strip.
test using a paper towel and quickly reinsert it into the meter
(within a period of two seconds).
 It will appear on the screen and the test results will not be saved in memory.
Do not remove the test strip from the meter until the display screen shows.
blank or otherwise the test results will not be cleared.

Urea
24 hours before the tests, the patient must follow a low-protein diet (meats
red) and fats.

Creatinine -- Albumin -- Protein Electrophoresis -- Proteins in 24 Hours and Clearance

of Creatinine:

The patient should avoid intense physical exercise for at least eight hours before the exam.
•Proteins in 24 hours and creatinine clearance require a 24-hour urine sample to
his evaluation. See 24-hour urine collection.

Bilirubin
Immediately after separating the serum, wrap the tube in aluminum foil. The light
this compound decreases rapidly.

9.2.4 Ionogram:
The puncture must be direct and without trauma.
Do not apply a tourniquet.
Traumatic punctures and hemolyzed sera increase values such as potassium.

10. SAMPLING FOR HEMATOLOGY:

The samples for Hematology are mainly conducted to assess cellular alterations.
such as anemias, leukemias, leukemoid reactions; the response to is also evaluated
hematopoietic treatments and infections and diseases that cause alteration of the
blood cells with similar clinical pictures.

For them, you must collect blood in a tube with EDTA for cell counts and Sodium Citrate.
for coagulation tests (with anticoagulant), it is enough to obtain a volume
Total blood volume approximately 10 ml for each of the tests. Blood must be drawn first.
the tube for coagulation tests and then for the complete blood count. The most common tests
what can be done with this sample include, among others:

Complete blood count Hemoparasites

Hemoglobin Hemoclassification.
Glycated Hemoglobin Sickle Cell Cycling Test.
Reticulocytes Hemoglobin Electrophoresis.
LE cells

The patient does not necessarily need to be fasting or in special conditions.

These tests can be performed at any time of the day.
Do not use the tourniquet for more than 30 seconds. See venipuncture.
Make a spread on a plate immediately after the puncture.
Gently mix to avoid clot formation.

10.1 COAGULATION TESTS:

10.1.1 Prothrombin Time (PT), Partial Thromboplastin Time (PTT), Fibrinogen.

Do not use a tourniquet.
Ask the patient what drugs they are taking (Coumadin, etc.). Write them down.
Use a tube with Sodium Citrate. (blue cap - Plasma)
Note existing bruises and petechiae.
Antihistamines, ascorbic acid, chlorpromazine, heparin, and salicylates prolong the
TPT.
The ingestion of alcohol, barbiturates, antibiotics, aminosalicylic acid, allopurinol,
TP levels increase, a diet rich in fats can reduce your level.

10.1.2 Coagulation time:

Make a direct and clean puncture with a 5 ml syringe for sample collection.
Extract three to five ml of blood, time the duration from the moment at
that the vein was penetrated.
•Distribute the extracted blood into three tubes without anticoagulant with approximately a
centimeter per tube, letting it fall down the walls.
•Immediately place in a water bath to incubate at 37 degrees Celsius; each
thirty seconds, observe the tubes until they coagulate completely.
The three individual times must be recorded and averaged to obtain the
coagulation time.
Normal values are between 5 to 15 minutes.

10.1.3 Bleeding time: Duke method (lancet):

Clean the earlobe with alcohol (if wearing earrings, they should be removed).
Hold the lobe of the ear for a few seconds, allowing its interior
contains a greater amount of blood.
With a new, sterile, and disposable lancet, make a puncture in the pressed area of the
ear lobe.
 Measure with a stopwatch or second hand the time it takes for the blood to clot.
originating from the small wound inflicted.
•Clean the bleeding every thirty (30) seconds. It should be done with filter paper taking
the drop of blood very superficially.
When the filter paper is no longer impregnated with blood, the clock or stopwatch is stopped and the
It records the time it took for the patient to coagulate from the moment of puncture.
until the moment the filter paper stops being saturated.

10.1.4 Bleeding time: Ivy method (with tensiometer):

Place the sphygmomanometer on the upper third of the arm at a pressure of 40 mm of
mercury (Hg).
Disinfect the inner part of the forearm where it is not very vascularized.
Make three quick punctures with a sterile lancet three centimeters apart.
Immediately start the timer and clean every thirty seconds with
filter paper blood but without touching the puncture site.
Stop the stopwatch when the bleeding stops. Note the time.
Normal values are 2 to 5 minutes.

10.2 CELLS LE:

The sample should be taken in a tube with anticoagulant (EDTA) and with glass beads.
to perform a defibrinization of the blood.

10.2.1 SAMPLE COLLECTION FOR HEMOPARASITES:

Total blood is used.

Clean the patient's fingertip with cotton soaked in alcohol, then puncture with
sterile lancet (or alternatively with a syringe needle).
Clean the first drop of blood with dry cotton and then take the next two drops.
placing at the ends of a previously marked glass plate.
With each of the drops and with the end of another plate, make in the shape of an N or zigzag.
extended dos of one centimeter by one centimeter or rectangular.
Let the spreads dry outdoors to color.
When the patient has received antimalarial drugs, it is recommended to take samples every six
hours, for three days.

11. SAMPLING FOR IMMUNOLOGICAL AND SEROLOGICAL TESTS

These tests evaluate different pathologies, they are a large number of exams that are
they can be performed on serological samples, but the most common ones are:
VDRL - Serology Seroagglutination for febrile patients
Rheumatoid factor (RA-Latex) Immunological pregnancy test (PIE)
Antistreptolysins (AELO- ASO) Blood classification.
C-Reactive Protein (CRP) FTA- ABSORBED- confirmatory VDRL.
HIV (AIDS) Hepatitis B Surface Antigen

It should be drawn into a tube without anticoagulant. (obtaining serum) a volume

approximately 10 ml for all tests, if there are many, two or more tubes will be taken.
The immunological pregnancy test can also be performed on a urine sample.

An eight-hour fast is recommended, but they can generally be done without

special conditions.
Do not consume fats or alcoholic beverages the day before.
The immunological pregnancy test (IPT) requires that the patient has a delay.
minimum of eight days during menstruation.
For HIV, the special information form that is required must be filled out.
legislation.

12. SAMPLING FOR MICROBIOLOGY AND MICROSCOPIC STUDIES

12.1 URINE FOR CYTOCHEMICAL OR URINE CULTURE:

The urine sample should be collected several hours after food intake.
The urine should be held for four (4) hours, preferably the first urine of the day.
tomorrow.
Wash the genitals with water and soap, retract the foreskin if male, separate the lips.
vaginal if she is a woman.
Discard the first part of the urine, from 15 to 30 ml.
Collect the following 50 to 100 ml directly in a sterile container, close immediately.
Take the urine to the laboratory as soon as possible or refrigerate it for no more than one hour.
The jar must be clean, preferably sterile, dry, with a screw cap, wide mouth and must be
well marked.
In children, a sterile plastic urine collection bag can be used. The bag will be placed
after washing the genitals by adhering it to the skin with an adhesive ring;
If it is not possible to collect the urine in the next 45 minutes, the bag must be changed.
for a new one.
Other methods of urine collection such as the use of catheters and suprapubic puncture.
They are reserved for the doctors to do.
12.2 24-HOUR URINE COLLECTION:

Urinate in the morning upon waking up and note the exact time (this urine should not be
collection).
Collect the subsequent urine samples (morning, afternoon, and night) in the
container provided by the laboratory, or in a clean spirits bottle. The
The container should preferably be opaque. Store the jar in the refrigerator.
collection (refrigerated).
The next day, exactly at the same time that the urine was disposed of the day before,
the last sample is collected.
Keep the jar during the study.
In case of forgetting to collect or partially or totally losing any portion of the urine,
collection for the study should be restarted.

The 24-hour urine sample is used for the following tests: Clearance of
Creatinine

12.3 BACTERIOLOGICAL OF VAGINAL FLUID:

12.3.1 INDICATIONS:

The patient should not perform vaginal washes or douches 24 hours before the examination.
The patient should not have used vaginal medications three (3) days before the
exam.
The patient should not have had sexual intercourse hours before the exam.
The procedure that will be performed will be explained to the patient.
The patient should not be menstruating.
It is suggested that the patient urinate before taking the sample.

12.3.2 VAGINAL FLUID COLLECTION:

Place the patient on a stretcher in a gynecological position where there is good

lighting for visualization, recommend the patient to relax for better
comfort and facilitate sample collection.
Using gloves, separate the lips to locate the vaginal opening, insert the speculum into
vertical form, rotate it until it is horizontal and insert it until you find the
cervix; open and fix the speculum to prevent it from closing.
Once the cervix is visualized, remove the excess cervical mucus with a gauze.
mounted on clamps or with an applicator; insert a sterile applicator into the canal
endocervical and gently rotate, wait a few seconds to allow for the absorption of the
germs to the applicator, remove the applicator and make the smear gently rotating the sample
to prevent the cell structures from breaking on the slide.
 Mark this sample as END (endocervix), this sample is important because it contains
We are going to look for intracellular gram-negative diplococci compatible with Neisseria.
Gonorrhea is important because women can be asymptomatic carriers of this bacteria.
Place the applicator in a tube with 0.85% saline solution.
•With a new applicator, take the sample from the pocket's background because that is where they are located.
microorganisms like Trichomonas and fungi; observe if erythema is present
characteristic of trichomonas infection. Place the sample on the slides and
mark as FS (dead end) and the applicator in the saline solution.
Take the pH of the flow with litmus paper directly when taking the flow with the last of the
applicators before placing it in the saline solution.
Remove the speculum and discard it in a bucket with sodium hypochlorite solution.
Let it air dry and fix in 95% alcohol as soon as possible to then color.
per gram.

12.4 URETHRAL SECRETIONS:

Using a dacron swab, take the sample directly from the urethra by dragging it well.
amount of sample. Spread on a clean slide while rotating to avoid damage.
cellular structures. Place the applicator in 0.85% saline solution.
Rotate again with another applicator and insert it into the saline solution.
Allow the sample to dry outdoors and gently fix with heat 2 to 3 times using
alcohol lighter.
Coloring by gram while controlling the staining.
If the secretion is scarce, perform a urine culture of the first portion of the urine.
in the morning.
12.5SPERMOGRAM:

12.5.1 INSTRUCTIONS:

The laboratory must always provide written instructions to avoid samples.

poorly gathered.
The container must be appropriate (wide-mouth), preferably sterile and without substances.
condoms.
The patient must maintain sexual abstinence for 2 to 5 days. There should not be more than seven.
days of sexual abstinence.
The patient must refrain from consuming alcoholic beverages for several days prior to the
exam.
The patient should not have had recent episodes of fever, nor undergone
surgical interventions nor prolonged pharmacological treatments; in any case the
the exam should be postponed for approximately one spermatogenic cycle, it is to say
seventy days.
The method of withdrawal cannot be accepted because of vaginal contamination.
produce a negative effect on sperm mobility and survival, in addition to
there is a risk of losing the first portion of the ejaculate in which there is a
greater number of active motile sperm.
Semen analyses cannot be repeated with less than seven days difference.
The ideal is for the sample to be collected in the privacy of a nearby facility.
laboratory; otherwise (at home) it must be sent to the laboratory before
an hour after the collection.
The patient must be referred to the Central Laboratory to take the sample.

12.5.2 SAMPLE TAKING:

The patient must urinate before taking the sample and then wash and rinse their hands well.
penis, in order to prevent bacterial contamination of the urethral portion.
• The sample must be obtained by masturbation and ejaculated into a container.
wide-mouth glass or plastic that is clean or preferably sterile, the container must
be warm to reduce the shock from the cold.
Condoms should not be used because they compromise the viability of sperm.
The method of interrupted coitus cannot be accepted because of vaginal contamination.
produce a negative effect on the mobility and survival of sperm, moreover
there is a risk of losing the first portion of the ejaculate in which there is a
higher number of active motile sperm.
Samples are not allowed to be analyzed if any part of the ejaculate has been lost.
The sample must be protected from extreme temperatures (below 20 degrees Celsius)
do not refrigerate, nor above 40 degrees Celsius.
 When delivering the sample, ensure it is clearly marked with the full name and
last names, the date and time of collection, and the period of sexual abstinence.

12.6 BACILIOSCOPY:

Investigation of acid-fast bacilli (AFB):

12.6.1 SPUTUM COLLECTION:

Take a fasting sample.

• Perform good oral hygiene and rinse with an antiseptic solution to prevent
contamination with the flora of the upper respiratory tract.
Cough strongly to detach the secretion from the bronchial tree and
lungs, deposit the sample in a wide-mouth sterile container.
It is recommended that the sample be the first of the morning and fasting to avoid.
food particles.
If the patient has difficulty expectorating, it is recommended that they collect the sample.
through "postural drainage": Place the base of the thorax on a pillow allowing the
arms and head towards the floor in order to facilitate the exit of bronchial secretions,
take a deep breath, hold the breath, and expel violently with a coughing effort.
It also facilitates the collection of the sample through the inhalation of water vapor that
it promotes the release of bronchial secretions.

For the urine sample, after cleaning the genitals, collect it in a container.
Sterile the last urine of the night and the first of the morning and those that are emitted in that period.

12.7 EOSINOPHILS IN NASAL MUCUS:

Material from both nasal cavities should always be collected separately, unless
The doctor wanted to study specifically one nostril.

12.7.1 PROCEDURE:

Preferably take the sample at the time of the rhinitic crisis.

Mark two containers preferably new, clean, dry, and free of grease with the
patient identification and right nasal cavity (RNC) and left nasal cavity (LNC).
Place a drop of Turk solution (glacial acetic acid plus methylene blue) of the
used for the count of leukocytes in each of the places where it will be
extend the sample.
 With a cotton swab, take a sample from each nostril trying to collect
abundant secretion, spread it well over the Turk solution avoiding any left over
clumps, as this makes it difficult to quantify the eosinophils.
If the sample is very dry, moisten the applicator in saline solution first.
to take the sample in order to try to drag a good amount.
Discard the applicators in a sodium hypochlorite disinfectant solution.
Leave to air dry before staining with Wright or Giemsa.

12.8KOH FOR FUNGUS STUDY:

The patient should not have used creams or topicals, talcum powder, deodorants, or nail polish.
nails 5 days before taking the sample, as they interfere with the exam.
The patient should not have had antifungal treatments 10 days prior to the examination.
Clean the sample collection area with a gauze soaked in sterile distilled water or
alcohol (do not use cotton).
•Perform a scraping with a sterile blade or scalpel of the edges of the lesion.
Take samples from different lesions.
Collect the scales in a sterile jar.
Always mark the samples with the anatomical site from which the sample was taken or
where is the injury located.
•

13. SAMPLING IN PARASITOLOGY

13.1 COPROLOGICAL, COPROANALYSIS, COPROGRAM:

Collect in the morning in a clean dry wide-mouth jar with a screw cap.
Do not mix with urine.
Do not take laxatives in case of constipation.
Never refrigerate the sample.
Serial coprological, collect three samples every other day.

13.2 HIDDEN BLOOD IN FECAL MATTER

The patient must have followed a diet without three days before the exam.
eating cold cuts, red meats, blood sausage, nor ferrous sulfate. The diet is
mandatory because the exam is performed to rule out gastrointestinal bleeding
produced by gastritis or ulcers.
13.3 TEST FOR PINWORMS:

Graham's technique or the tape method.

The patient should not have bathed.
Place the tape (in tube form) in the perianal area of the patient.
The adhesive tape can be applied by the patient at night and removed in the
Tomorrow, as the parasites come out to lay their eggs at night.
Place the tape on a slide pushing it down firmly.

14. SAMPLE COLLECTION FOR MICROBIOLOGY (CULTURES).

14.1 Urine Cultures

First morning urine.

•Preliminary instructions and delivery of manuals and sterile vial.
Sow immediately upon receiving the sample in UROBACTER after shaking. See usage of
UROBACTER in the procedures manual.
Always note the patient's age and sex.
Presence or absence of pregnancy and the months of gestation.
•Note whether it is control or not.

14.2 COCULTURES:

It is not necessary to transport in buffered glycerin.

Normal sample as for coprological.
•Record the patient's age and drug treatments.

14.3 CULTURE FOR GONCOCCUS - THAYER MARTIN :

• Follow instructions for taking urethral secretion samples for men and flow.
vaginal for women.
•Sow directly on the sample collection site. See use of GONOBACTER in
procedures manual.
With a sterile swab, take a sample from the exocervix and culture it in Thayer-Martin medium.
T.M.).
With a sterile swab, take part of the urethral secretion or make a smear of the lesion and
sow in the middle Thayer Martín (T.M.)
14.4 ABSCESSES:

Samples of abscesses from any part of the body must be taken by the
bacteriologist or assistants trained directly in the laboratory, to be inoculated
immediately in the appropriate culture media thus avoiding contamination of the
samples due to excessive handling and transport ensuring a truthful result.

Puncture the abscess with a sterile needle and syringe.

If the abscess has previously burst, collect the purulent material using
sterile spatula or sterile bacterial loop.
Place the material in a sterile glass container with a lid or leave the obtained material in the
syringe and place the rubber cap on the needle to prevent air entry or contamination
through the needle (in case of sample referral).
•Carry out the necessary extensions for coloring and direct examination and allocate part of the
material for crops, if needed.
Process the samples within two (2) hours.

14.5OTHER SAMPLES:

There are other samples that can be processed in the laboratory, their collection or
This procedure is exclusively and strictly reserved for specialist medical personnel.
Generally, these specimens are managed in a hospital setting, but occasionally they may be
processed at an individual level, among these we have:

Bronchoalveolar lavage.
Cerebrospinal fluid.
Bronchial Secretions.
Suprapubic Puncture.
Use of Catheters.
Ocular Material.
Pleural, pericardial, peritoneal, joint or synovial fluid.
Gastric Lavage.
Biopsies.
Bone Marrow.
Paranasal Sinuses and Pharynx.
Blood cultures.
Prostatic Fluid.
 These samples are taken and processed for the detection of pathogens such as
viruses, fungi, parasites, and bacteria; must be collected in very clean or sterile places
and under special aseptic conditions to prevent the patient from becoming contaminated or complicated
due to infections introduced during the sampling.
They are samples that must be transported immediately to the laboratory for analysis.
in a very sterile manner, they must be handled by professional or trained personnel to
Hello. Your process should be completed within two hours.
If any type of analysis of these is required, the assistant must call the laboratory.
before the sample is taken, to prepare the elements, culture media, and reagents
necessary for its processing.

14.6 WATER AND FOOD CROPS:

These samples must be taken by trained personnel and sent at least
possible time to the laboratory for processing, the assistant or responsible person must
communicate in advance with the Laboratory.

15. SAMPLING FOR CERVICAL VAGINAL CYTOLOGY

15.1 INTERVIEW:

Allow the patient to have privacy and assure her that the provided information
they are confidential for her.
Inform him about the meaning of cytology, treating it very delicately, with respect and
courtesy.
Encourage her to claim the result in a timely manner.
Do not take the cytology if the patient has applied intravaginal medications three days or
less than before the sample collection and/or if you have had sexual relations in the last 24-48 hours
previous.
Explanation of the procedure to the patient.
Complete the form to request the cervical-uterine cytology:
a) Time since the last cytology was performed.
b) Full names.
c) Age.
d) Pregnancies, births, abortions.
e) Date of the last menstrual period. If menopausal, date of this.
f) Planning or contraception method.
g) Neck treatments: Cauterization, Conization, Radiotherapy, Hysterectomy (causes
of this and what treatments), others and date.
h)Síntomas actuales: Ninguno, Flujo anormal, Dolor pélvico, sangrado post-coito y post-
menopause, burning, itching, others.
Label the sheet with the medical record number, identity document number, and/or
patient's name.
15.2 PRECAUTIONS:

Do not perform bimanual exploration before taking the cervico-uterine cytology sample.
Use the speculum without lubricants; if necessary, it can be moistened with distilled water or
sterile water, never with saline solution, this inflames the cells causing us harm
diagnostic interpretation.
Do not use hairspray as a fixer, as it alters the sample at the moment of the
coloration in the laboratory.
Do not leave the sample to dry outdoors or where it is exposed to strong winds; the items
environmental cell phones will be of such magnitude that they will prevent the proper reading of
smears, making it unfit for diagnosis.

15.3 PROCEDURE:

Initially, the patient should be asked to urinate before positioning her.

gynecological for the exam, provide kind treatment to the patient.
We must have the plate and the order clearly marked.
The patient is placed in a gynecological position to avoid unnecessary exposure. It should be
to be as comfortable as possible.
With sterile gloves, separate the labia majora, carefully observing them to see
if pathological changes are found (Bartholin cysts, condylomas, warts in
perineum that also simulate condylomas), and inform them in the medical history sheet.
The speculum is introduced (without lubrication) vertically, slowly turning it to the
horizontal position, it opens gently to reveal the neck.
Visualizing the neck, secure the speculum in the open position and screw it in.
•Observe characteristics of the vagina and cervix; identify the sites for taking a cytology sample; if
there is mucus, purulent discharge or blood on the surface of the neck, it should be removed
gently with a cotton swab, gauze moistened with sterile water or distilled water
before the sample is taken.
After removing the mucus or exudate from the neck, one proceeds to observe.
Characteristics of the neck: if it is believed to be eroded, carefully loosen the screw of the
speculum trying to keep the neck in a normal position (like closed), if after this
the process does not show reddish lesion around the endocervical opening, we say that this is
Yes, if it is the opposite, it is reported as eroded, being very careful to know it.
differentiate from an ECTROPION.

If the cytology is functional, three samples are taken from the same slide proceeding as follows:

a) Vaginal Sample: Scrape both walls (upper lateral fundus) with a spatula or
apply the tongue previously moistened with distilled or sterile water. Carry out the spreading on
horizontal, uniform, and thin at the edge of the sheet. This sample will serve to
observe infectious processes (Gardnerella, Trichomonas, Fungi, etc.).
b) Exocervical Sample: Take the sample from the squamocolumnar junction in the same way with
a spatula or tongue depressor previously moistened with distilled or sterile water making it
rotate 360 degrees with firm pressure (scraping epithelium, do not forget that the cytological sample is
abrasive, it is not the flow that matters but the epithelium), taking care not to touch the walls of
the vagina to avoid contamination with cells from the lower portion of the walls
vaginal or from the bottom of the lateral pouches (Douglas pouch), perform an extension in
horizontal, uniform, and thin in the center of the sheet. This sample is fundamental.
to detect Cervical Cancer, as it is in the Squamocolumnar junction where it starts
Cervical cancer.

c) Endocervical sample: With a cytobrush previously moistened with distilled water or

gently introduce the sterile instrument into the endocervical canal, up to the point where it meets
an obstacle or where the bristles end, proceed to rotate it 360 degrees with firm pressure
trying to touch the endocervical walls; remove it gently, perform the extension in
the left end next to the number, in a uniform and thin horizontal or circular shape,
turning the cytobrush in a circular motion (360 degrees) over the slide, in such a way that the epithelium
endocervical remains adhered to it. In oncological cytology, only 2 samples are taken:
Exocervical and Endocervical. Through this sample, we will be able to detect an Adenocarcinoma.
or alterations specifically in endocervical cells, which are of the Columnar type.
Cytologically, we realize the quality of the sample by being able to observe them.
cells.
 Fix the slide immediately after taking the sample using one of the
following techniques:

a) Atomizer (Citospray), contains propylene glycol, should be used at a distance

approximately 20 to 20 cm. between the sheet and the atomizer, dispersing evenly
and thin avoiding leaving a thick film that helps wash the plate and thus alter the
cytological diagnosis; subsequently let it dry in the environment for 5 minutes, this
the method is suitable for cytologies that are going to be sent to other laboratories;
previously wrapped in each corresponding story, they proceed to be packed and
labeled for shipping.
b) Liquid fixative, it is 95% ethyl alcohol. Once the cytology is taken, the slide is submerged.
in the container with alcohol for 15 minutes until the coloring process begins.
There is also another fixative that is a mixture of 95% ethanol and polyethylene glycol;
Once the cytology is taken, 4 or 5 drops of the fixative should be applied with a dropper.
surface of the sheet and with tilting and rocking movements scatter the drops over the
surface and let dry in the environment. This method is also used for transporting
the cytological samples to the laboratories in the same form as the Citospray fixative.

Unscrew the speculum and gently remove it from the vagina.

•Record the characteristics of the neck: Healthy, Eroded, Ectropion, Congestive, Polyps,
Naboth cysts, leukoplakia, erythroplasia, atrophic, condylomas, etc.
Record the characteristics of the vagina: Condylomas, Gardner cysts, etc.
•Register the characteristics of the Vulva: Leucoplasia, Bartholin's cysts, Condyloma
pointed, Herpes, etc.
Finally, the cleaning is carried out by discarding the material and equipment used.
sample in accordance with waste disposal precautions and biosafety
known and required.

Note:
If abundant vaginal discharge is observed during the exam, a Direct test should be taken first.
gram of vaginal discharge, then clean the discharge very well with an applicator and proceed to take the
sample.

16. SAMPLING FOR SPECIALIZED TESTS

They are generally serum or plasma samples using the anticoagulant EDTA or
heparin that must be promptly separated from red cells, protected from the
exposure to light, preferably without hemolysis and stored in refrigeration or freezing
to avoid the loss or deterioration of the components to be analyzed.
The samples do not require prior fasting from the patient, so they can be
taken at any time but if possible lipemic samples that produce are avoided
Some interferences. The minimum volume is one milliliter (1 ml) for its analysis.
Hemolyzed, icteric, lipemic, or contaminated samples are generally
inadequate for these tests.
 1. Thyroid Profile (T-3, T-4, TSH).
2. Chorionic Gonadotropin (B-HCG) qualitative and quantitative.
3. Prolactin, preferably a sample at ten in the morning on complete fasting.
Luteinizing Hormone (LH).
5. Follicle Stimulating Hormone (FSH).
6. Antinuclear Antibodies (ANA).
[Link]:Ig E , Ig M , Ig G, Ig A totales cada una.
8. Immunoglobulins Ig G and Ig M (Rubella - Toxoplasmosis - Cytomegalovirus CMV).
[Link]ígenos para cáncer:ACE, CA 125, Ca 15-3, AEP,
[Link]:Carbamazepina, Digoxina, Litio, Teofilina, Ciclosporina, Fenitoina, Primidona.
[Link]:B12, A, D, C., Acido Folico.
[Link] G or Ig M antibodies against: ANA, Antimycoplasma, HIV, Hepatitis A, Hepatitis B
Hepatitis C, Antimicrosomal, Toxoplasma, Rubella, Antithyroglobulin
[Link]:Androstenodiona, Antidiuretica ADH (Vasopresina), del Crecimiento (GH),
Insulin,
[Link] Tests.
[Link]: Aldosterone, Amylase in serum or urine (urine collected over two hours)
is sufficient), GOT (AST), GPT (ALT), Total creatine phosphokinase and MB, Cholinesterase,
Gamma Glutamyl Transpeptidase, Lipase,

• Bleed in a dry tube without anticoagulant, 10 ml for all tests. (serum).

Preferably an 8-hour fasting, but it can be taken at any time.
These serum samples must not be hemolyzed.

THE 24-HOUR URINE SAMPLE is used to perform the following tests:

Creatinine Clearance. Albumin. Free cortisol in urine.
Delta Aminolevulinic Acid (ALA). Catecholamines Football.
Vanilmandelic Acid (VMA) Chlorine. Cortisol.
5-Hydroxyindoleacetic Acid (5-HIAA) Aldosterone.
Phenylketonuria (PKU). Sodium. Proteins.
Creatinine Porphyrins. Testosterone.
Fractionated estrogens. Pregnadiol. Potassium
Follicle Stimulating Hormone (FSH)
Neutral cetosteroids (17 cetosteroids, 17 hydrox corticosteroids)
This sample must always be refrigerated during its collection. See urine collection of
24 hours.

Vanillylmandelic Acid (VMA) should preferably be collected in containers.

plastics and the urine acidified with 20 milliliters of 6 mol/L HCl solution; and it should be avoided the
ingestion 48 hours before Banana, Coffee, Tea, Chocolate, Vanilla, and Plantains.

5-Hydroxyindoleacetic acid (5-HIAA) must be collected with 15 milliliters of solution.

6mol/L of HCl or 15 c.c of glacial acetic acid at 33%, storing it in refrigeration.
 Aldosterone and Catecholamines, Estriol, with the addition of 15 grams of acid
boric

Cortisol, with 20 c.c. of 33% glacial acetic acid.

HEPARINIZED TUBES:

It is used in some routine studies as well as specialized ones. Its presentation can
include sodium or lithium concentrations. In general, heparin with lithium is used for
Chemical studies and sodium heparin is used for lymphocyte study (e.g. HLA).
Also for samples such as Adrenocorticotropic Hormone (ACTH), T4 Lymphocytes and
T8, Ammonium, HLA-B27 Antigen (Human Lymphocyte B27), Karyotype, LE Cells with pearls of
glass, Arterial Gases,

17. TRANSPORT AND SHIPPING OF SAMPLES

Clean the containers well to mark them and thus avoid the peeling off of the labels.
adhesives.
Do not mark samples on the lids, always do it on the containers.
• Mark with names and surnames, reception code and age.
Wrap the plates in tissue paper and always place them in a plastic bag.
Do not allow the plates to be placed in the transport refrigerator.
Send urine and stool samples separately and in plastic bags. Tighten the caps properly.
Separate tubes with serums, blood tests, and clots to expedite receipt.
samples, the plastic bags must be red.
Mark each of the bags.
Always remind the transporter that liquid samples must be upright.
• Record all samples on the remittance sheet, by patient, analysis, ID, number
internal if applicable, age, and delivery time, etc.
• Check that the transport cooler has a rack and a thermometer.

20. SENDING OF ONCOLOGY SAMPLES (FLOW CYTOMETRIES, BIOPSIES,

KARYOTYPES, MYELOGRAMS, SPECIAL STAINING E
IMMUNOHISTOCHEMISTRY

These are considered special samples, due to the characteristics of the sample, for
being invasive in its takeover and by being referred to special entities.
Samples are only taken and received in the laboratory from Monday to Thursday. This is because they must be
processed the same day and read the next day, and it could not be done on the weekend.
They are taken by oncologists and hematologists, who take a purple topped tube to
immunological classification of leukemia, a tube or bottle with Bown reagent or preservative
for the biopsy, and several slides fixed at room temperature for the myelograms and
special colorations and immunohistochemistry.
They are transported immediately to the laboratory at room temperature and must be stored as such.
up to 24 hours, or at refrigeration temperature for a maximum of 48 hours.

The orders are photocopied, the original and a copy are given to the messenger to bring back the copy.
signed for receipt.
The bone marrow smears, bone marrow biopsies, immunological classification cytometries of leukemia and
of stem cells, special stainings and immunohistochemistry are referred to the HPTU.
The karyotypes to the Cytogenetics laboratory of UdeA CIU.
The HTLA for compatibility study at the Cytometry and Genetics laboratory of UdeA
CIU.
These results are delivered electronically to the blood bank's email and the address of the
laboratory.
 [Link]

1. Yen, Samuel S. C. Jaffe, Robert B. Reproductive Endocrinology: Physiology,

Pathophysiology and Clinical Management. Third Edition. Pan American Medical Publishing. Buenos
Aires (Argentina), 1993. p 977.

[Link]-Sanford-Davisohn. Clinical Diagnosis and Treatment by the Laboratory. Eighth

Edition. Salvat Editores, Barcelona (Spain), 1988. p 613-623.

Clinical Interpretation of the Laboratory

Panamericana, Bogotá, 1993. p. 387.

Miale, John B. Hematology. Laboratory Medicine. Sixth edition. Reverté Publishing, S.A.
Barcelona, 1985.
5. Eduardo, Gómez G. Alberto Gómez G. Manual for Sample Collection in the
Clinical Laboratory. Laboratory and Medicine. First edition. Diseditorial publishing, Santafé of
Bogotá, 1996.
6. Angel M, Gilberto. Clinical Laboratory Dictionary. First Edition, Medical Publishing
International. Bogotá, Colombia, 1996.