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 Important notes
Thank you for purchasing the RTP® Pathogen Kit from Invitek Diagnostics.
The product is intended for the manual isolation of nucleic acids (bacterial DNA, viral
DNA/RNA) from a variety of clinical samples using Spin Column technology.

WARNING! Improper handling and use for purposes other than those intended may cause
danger and damage. Therefore, we ask you to read through these instructions for use carefully
and follow them precisely. Always keep them accessible. To avoid personal injury, also observe
the safety instructions.
All versions of the instructions for use can be found on our website for download from the
following address: [Link]

Technical support:
techsupport@[Link]

GERMANY
Haynauer Str. 60, 12249 Berlin, Germany

PORTUGAL
Zona Industrial de Tondela, ZIM II, Lote 6, 3460-070 Tondela, Portugal
+351 232 817 817

© 2025 Invitek Diagnostics, all rights reserved.

The kit complies with REGULATION (EU) 2017/746 on in vitro diagnostic medical devices. However, it is not intended
for in vitro diagnostic use in countries where REGULATION (EU) 2017/746 is not recognised.

Trademarks: InviSorb®, PSP®, InviMag®. Trademarks, commercial brands, etc. mentioned in this document, even if not
specifically identified as such, should not be considered unprotected by law.

InviGenius®, InviMag®, InviSorb®, Invitek®, InviTrap®, MSB®, PSP®, RTP® are registered trademarks of Invitek
Molecular GmbH.
Table of Contents
1. Product information ........................................................................................................ 3
1.1 Kit contents .................................................................................................................. 3
1.2 Symbols used on product and labelling ........................................................................ 3
1.3 Reagents and equipment to be supplied by user.......................................................... 4
1.4 Storage, appearance, and shelf life .............................................................................. 4
1.5 Intended use ................................................................................................................ 5
1.6 Performance characteristics ......................................................................................... 5
1.6.1 Performance data by matrix ......................................................................................... 5
1.7 Interfering substances .................................................................................................. 7
1.8 Principle and procedure ............................................................................................... 8
2. Nucleic acid extraction with the RTP® Pathogen Kit ............................................................. 8
2.1 Before starting a protocol ............................................................................................. 8
2.2 Sampling and storage of starting material .................................................................... 9
2.3 Preparation of starting materials ................................................................................ 11
2.3.1 Serum, plasma, other cell-free body liquids................................................................ 11
2.3.2 Swabs ........................................................................................................................ 11
2.3.3 Stool samples (supernatant) ...................................................................................... 11
2.3.4 Cultivated bacteria ..................................................................................................... 11
2.3.5 Urine 12
2.3.6 Respiratory samples: tracheal, bronchoalveolar lavage (BAL) and sputum ................ 12
2.3.7 Tissue Biopsies .......................................................................................................... 12
2.3.8 Cell culture supernatants ........................................................................................... 12
2.4 Short protocol RTP® Pathogen Kit ............................................................................. 13
2.5 Protocol for simultaneous isolation of bacterial DNA and viral DNA/RNA from liquid
samples ..................................................................................................................... 14
3. Warranty...................................................................................................................... 15
4. Safety instructions ......................................................................................................... 15
4.1 Reagent Specifications and Hazard Classification ..................................................... 16
4.2 List of Hazard Warnings and Precautionary Statements ............................................ 17
5. Appendix ..................................................................................................................... 18
5.1 Troubleshooting ......................................................................................................... 18
5.2 Other documents and further information ..................................................................... 19
5.3 Ordering information .................................................................................................. 19
 1. Product information
1.1 Kit contents

50 preparations 250 preparations

Catalogue No. 1040500200 1040500300
Extraction Tubes 50 tubes 250 tubes
Resuspension Buffer R 30 ml/bottle 150 ml/bottle
Binding Solution empty bottle empty bottle
(fill with 99.7% Isopropanol) (Final volume 30 ml) (Final volume 120 ml)
Wash Buffer R1 20 ml/bottle 80 ml/bottle
12 ml/bottle 50 ml/bottle
Wash Buffer R2
(Final volume 60 ml) (Final volume 250 ml)
Elution Buffer R 15 ml/bottle 60 ml/bottle
RTA Spin Filter Set 50 sets 250 sets
RTA Receiver Tubes 150 pieces 750 pieces
1.5 ml Receiver Tubes 50 pieces 250 pieces
Short Protocol 1 leaflet 1 leaflet

1.2 Symbols used on product and labelling

Manufacturer

Lot number
Unique identifier of a medical device
Catalogue number
Expiry date
Consult Instructions for use
Temperature limitation
Do not reuse
Amount of sample preparations
in vitro diagnostic medical device

CE marking

RTP® Pathogen Kit 3 DE 806.02_EN

 1.3 Reagents and equipment to be supplied by user
Lab equipment:

x Microcentrifuge
x Optional: centrifuge for 15 or 50 ml
x Thermo shaker (37°C - 95°C)
x Measuring cylinder (250 ml)
x Disposable gloves
x Pipette and pipette tips
x Vortex mixer
x Reaction tubes (1.5 ml, 2.0 ml)

Liquids and solvents:

x DNase/RNase free water or 1 x PBS to adjust sample volume
x 96 - 100 % ethanol (non-denatured)
x Isopropanol
x Optional (for respiratory samples with high viscosity): saturated acetylcysteine
(ACC) solution (200 mg/ml)

1.4 Storage, appearance, and shelf life

Shelf life: All buffers and kit components should be stored at room temperature and have a shelf
life as indicated on the outer kit package label.
After opening, individual components of the kit, as well as components prepared accordingly
before first use, have a shelf life of 3 months.
Before each use, make sure that all components are at room temperature. If there are
temperature-related precipitates in the solutions, dissolve them by carefully warming (up to 30°C).
Room temperature (RT) is defined as a range from 15-30°C.
Wash Buffer R1 and Wash Buffer R2: after adding ethanol, they should be firmly closed and
stored at room temperature.
Binding Solution: after adding isopropanol, it should be firmly closed and stored at room
temperature.

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 1.5 Intended use
The RTP® Pathogen Kit is a Spin Column technology based nucleic acid extraction kit, intended
for the simultaneous isolation and purification of bacterial DNA and viral DNA/RNA.
The kit can be used for a variety of human sample types, such as serum and plasma (from blood
stabilized in EDTA or citrate but not heparin), respiratory samples, supernatant from stool
suspension, cultivated bacteria, cell culture supernatants, biopsy material/tissue, urine, and other
cell-free body fluids.

1.6 Performance characteristics

The performance evaluation of the RTP® Pathogen Kit was performed by extracting nucleic
acids from various human clinical samples, including plasma, blood, nasopharyngeal swabs,
faeces, urine, and respiratory samples. These matrices were contaminated with different amounts
of bacterial and viral pathogens. Where available, clinical samples from individuals with confirmed
diagnoses of various diseases were used.
The extraction of pathogen nucleic acids (bacterial DNA, viral DNA and viral RNA) was performed
according to the procedures described in this document. The efficiency of the extraction was
evaluated by detecting the target nucleic acids by qPCR (for DNA) or RT-qPCR (for RNA).

1.6.1 Performance data by matrix

Serum and plasma
The RTP® Pathogen Kit demonstrated high sensitivity for HBV detection in low-titre serum
samples, which is crucial for clinical monitoring and early diagnosis. In assays with human
cytomegalovirus (hCMV) subjected to serial dilutions in plasma samples, the expected
relationship between decreased viral concentration and increased Ct values was observed,
demonstrating the quantitative accuracy of the method. This linear response confirms the
applicability of the kit in viral quantification and monitoring of viral loads in patients.
Urine
Urine is a matrix commonly considered challenging due to the presence of salts and compounds
that can inhibit PCR reactions. Nevertheless, the RTP® Pathogen Kit demonstrated consistent
ability to extract and detect Chlamydia trachomatis DNA, with uniform and reproducible Ct values.
This highlights the efficiency of the process in naturally inhibitory matrices, making it suitable for
use in the diagnosis of urogenital infections.
Faeces (supernatant)
Faeces are a highly complex matrix containing polysaccharides, bile acids and cell debris, which
can impair nucleic acid extraction and amplification. However, the kit ensured reliable detection
of Clostridium difficile, demonstrating resistance to inhibitors. This robustness reinforces the
method's potential for applications in the diagnosis of enteropathies and epidemiological studies,
where sensitivity is critical.
Nasopharyngeal swabs
Clinical samples subjected to extraction with the RTP® Pathogen Kit confirmed the detection of
SARS-CoV-2 in positive cases and the absence of amplification in negative cases, demonstrating
high diagnostic specificity. The quality of the extraction allowed clear detection of viral RNA, even
in moderate viral loads. This performance confirms the applicability of the kit in respiratory
surveillance testing and screening for emerging viral infections.

RTP® Pathogen Kit 5 DE 806.02_EN

 Cultured bacteria
Bacillus subtilis colonies were used as a model for Gram-positive bacteria, characterised by a
thick cell wall that often hinders lysis and DNA extraction. The RTP® Pathogen Kit demonstrated
high efficiency, providing high-purity DNA (A260/A280 ratio: 2.13 ± 0.02) and excellent yield.
Respiratory samples
Sputum samples from individuals diagnosed with Mycobacterium tuberculosis, Metapneumovirus
and Mycoplasma pneumophila were successfully processed. The detection of different pathogens
(bacterial DNA and viral RNA) demonstrates the versatility of the kit in different types of infectious
agents.
Tissue samples
The RTP® Pathogen Kit was applied to biopsies containing Human Papillomavirus (HPV), a
particularly difficult matrix due to the presence of structural proteins, lipids, and cellular
components that can interfere with extraction. Performance was consistent, with a wide Ct range
compatible with different viral loads present in clinical samples. This demonstrates the kit's ability
to handle solid clinical material, providing DNA suitable for molecular analysis and
histopathological studies.

Table 1 . Performance of the RTP® Pathogen Kit in different matrices

Mean Ct ± SD |
Matrix Pathogens/Target Target Type Condition
Yield (µg)
Human serum HBV Viral DNA Low viral titre Ct 35.14 ± 0.82
Undiluted Ct 26.59 ± 0.3
Human
Human plasma cytomegalovirus Viral DNA Dilution: 10-1 Ct 29.05 ± 0.44
(hCMV)
Dilution: 10-2 Ct 33.32 ± 1.39
Contaminated
Urine Chlamydia trachomatis Bacterial DNA Ct 32.75 ± 1.92
human urine
Contaminated
Faeces Clostridium difficile Bacterial DNA Ct 24.81 ± 1.19
human faeces
Nasopharyngeal Contaminated
SARS-CoV-2 Viral DNA Ct 33.24 ± 0.64
swabs swabs
Bacterial
Cultured bacteria Bacillus subtilis Bacterial DNA 16.13 ± 2.20 µg
cultures
Metapneumovirus
Respiratory Contaminated
Mycoplasma Bacterial DNA 28.00 ± 3.185
samples sputum
pneumophila

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 1.7 Interfering substances
The impact of potential interfering substances on the efficiency of nucleic acid extraction was
evaluated using artificially contaminated samples. The substances were added at concentrations
representative of the highest levels reported in the sample material.
Relevant substances were tested at the calculated maximum concentrations (Table 2). Extraction
was performed according to the established protocol, with duplicates tested for each condition.
The efficiency of nucleic acid recovery was evaluated by RT-qPCR for viral RNA (InviScreen®
IBV Detection Kit | Item No.: 6016002200) and by qPCR for bacterial DNA (InviScreen®
Salmonella spp. Detection Kit | Item No.: 6015007200). For bacterial DNA and mammalian
DNA, the evaluation was performed by spectrophotometric measurement and gel analysis. For
RNA, photometric measurement, gel analysis, and RT-qPCR were used. The results were
compared with control samples processed identically but without interfering substances.

Table 2. Potentially Interfering Substances in RTP® Pathogen

Potential Tested
Active ingredient(s) Target(s) Interference
Interferent concentration
Ethylenediaminetetraacetic
EDTA 10 mg/ml IBV | Salmonella NO
acid
Haemoglobin Haemoglobin 7.5 mg/ml IBV | Salmonella NO
Cholesterol Cholesterol 10 mg/ml IBV | Salmonella NO
Fatty acids Gamma-linolenic acid 0.1 mg/ml IBV | Salmonella NO
Albumin Serum albumin 40 mg/ml IBV | Salmonella NO
Polysaccharides Starch 100 mg/ml IBV | Salmonella NO
Urea Urea 50 mg/ml IBV | Salmonella NO
Alcohol Ethanol 3 mg/ml IBV | Salmonella NO
Salts Sodium chloride 7 mg/ml IBV | Salmonella NO
Ampicillin Ampicillin 100 µg/ml IBV | Salmonella NO
Colistin Colistin 50 µg/ml IBV | Salmonella NO
Nalidixic acid Nalidixic acid 5 µg/ml IBV | Salmonella NO
Anti-inflammatory Ibuprofen 4 mg/ml IBV | Salmonella NO
Heparin Lithium heparin 0.2 mg/ml IBV | Salmonella YES
IBV: Infectious Bronchitis Virus.

Negative interference was observed with high concentrations of heparin, evidenced by increased
CT values or failures in PCR amplification.
All other substances tested did not compromise nucleic acid extraction or subsequent detection,
confirming the robustness of the extraction protocol.

RTP® Pathogen Kit 7 DE 806.02_EN

 1.8 Principle and procedure
I. Sample lysis
The kit contains Extraction Tubes which contain a lyophilized mix of Carrier RNA,
Proteinase K, lytic enzymes and Lysis Buffer to perform a 1-step sample lysis. To start the
extraction procedure simply add the sample to the Extraction Tube.
Samples are lysed at different elevated temperatures while continuously shaking.
II. Binding of nucleic acids
By adding Binding Solution to the lysate, optimal binding conditions are adjusted. Each
lysate is then applied to an RTA Spin Filter and nucleic acids are adsorbed to the
membrane.
III. Washing to remove residual contaminants
Contaminants are efficiently washed away using Wash Buffer R1 and Wash Buffer R2,
while nucleic acids remain bound to the membrane.
IV. Elution of nucleic acids
Nucleic acids are eluted from the RTA Spin Filter using 60 - 200 µl Elution Buffer R.

2. Nucleic acid extraction with the RTP® Pathogen Kit

2.1 Before starting a protocol
When using the kit for the first time make sure all buffers and reagents are prepared as
indicated:
Buffer preparations prior first use: 50 preparations
Binding Solution (empty bottle): Fill 30 ml 99.7% isopropanol (molecular biology grade)
into the bottle, always keep the bottle firmly closed.
Wash Buffer R1: Add 20 ml of 96 -100% ethanol to the bottle. Mix thoroughly, always keep
the bottle firmly closed.
Wash Buffer R2: Add 48 ml of 96 -100% ethanol to the bottle. Mix thoroughly, always keep
the bottle firmly closed.
Buffer preparations prior first use: 250 preparations
Binding Solution (empty bottle): Fill 120 ml 99.7% isopropanol (molecular biologic
grade) into the bottle, always keep the bottle firmly closed.
Wash Buffer R1: Add 80 ml of 96 -100% ethanol to the bottle. Mix thoroughly, always keep
the bottle firmly closed.
Wash Buffer R2: Add 200 ml of 96 -100% ethanol to the bottle. Mix thoroughly, always keep
the bottle firmly closed.

x Adjust the thermo shaker to 37°C.

x Adjust thermo shaker/heating blocks to 65°C and 95°C
x Warm up the needed amount of Elution Buffer R to 65°C (60 - 200 µl Elution Buffer R are
needed per sample).
x Determine the number of required reactions including controls and label the needed amount
of RTA Spin Filters (lid) and the needed amount of 1.5 ml Receiver Tubes (per sample:
1 Receiver Tube is needed).

RTP® Pathogen Kit 8 DE 806.02_EN

Extraction control
Refer to the manufacturer’s instructions to determine the optimal amount of extraction control for
specific downstream applications.
Extraction control DNA or RNA should be added to the lysate after the heating step. For optimal
purification efficiency, extraction control molecules should be longer than 100 nucleotides, as
smaller molecules are not efficiently recovered.

2.2 Sampling and storage of starting material

The manufacturer is not responsible for sample collection, but only provides recommendations on
its storage and appropriate conditions, as validated during device development.
Storing samples correctly is essential to ensure reproducible and high yields. Yields may vary
depending on factors such as donor health, age and sample type, transport conditions and storage.
To preserve the integrity of the samples, the following storage conditions are recommended:
x Short term (up to 2 hours): keep samples on ice or under controlled refrigeration between
2°C and 8°C.
x Up to 24 hours: store at -20°C
x Long-term storage: store at -80 °C, preferably in aliquots to avoid repeated freeze-thaw
cycles.

Repeated freeze-thaw cycles of samples should be avoided to prevent nucleic acid degradation.
In general, the best results are obtained with fresh samples. It is recommended that technical
guidelines, such as CEN/TS and ISO standards, on the pre-examination process for molecular
diagnostics under the European In Vitro Diagnostic Medical Devices Regulation (IVDR) be taken
into account.
The following table provides specific recommendations for collection, preparation, and storage for
each sample category, in accordance with the applicable technical requirements.

RTP® Pathogen Kit 9 DE 806.02_EN

 Sample type Sampling Recommendations Critical observations
Do not vortex whole blood to prevent haemolysis.
Venous blood with EDTA or citrate Separate plasma/serum within 12 hours after Avoid freeze-thaw cycles to prevent protein denaturation/precipitation.
anticoagulant (not compatible with collection. In the presence of cryoprecipitates, centrifuge at 6,800 x g for 3 minutes
heparin) and use the supernatant immediately.
Transfer supernatant to new tubes if obtained in
systems without gel separators.
Homogenise well before use. Avoid freeze-thaw cycles to prevent protein denaturation/precipitation.
Serum, plasma, and other acellular body
fluids Allow serum WXEHVWRVWDQGIRU•PLQXWHVEHIRUH In the presence of cryoprecipitates, centrifuge at 6.800 x g for 3 minutes
centrifugation. and use the supernatant immediately.
Dry swabs Store in a dry place at 4-8°C. Do not freeze.
The stabilising liquid can be handled as a cell-free
body fluid. It is recommended to check the compatibility of the components before
Swabs with stabilising medium
use. Certain stabilising agents may compromise the efficiency of the kit.
Store according to the manufacturer's requirements.
Contains DNases and RNases that can cause rapid degradation of
Faeces Freeze immediately at -80°C.
DNA/RNA.
Sediment after cultivation and freeze at -20°C or -
Cultured bacteria
80°C for long-term storage.
An initial volume of 15-50 ml of urine is
recommended.
Urine Contamination with urea may inhibit PCR reactions.
Centrifuge to sediment bacteria and remove
supernatant.
Tracheal secretion, bronchoalveolar Contains DNases and RNases that can cause rapid degradation of
Freeze immediately at -80°C.
lavage (BAL) and sputum DNA/RNA.
Freeze immediately at -20°C or -80°C.
Tissue biopsies Avoid freeze-thaw cycles.
Thaw the sample in the lysis mixture.

RTP® Pathogen Kit 10 DE 806.02_EN

 2.3 Preparation of starting materials
In the following the preparation of the sample lysis for different starting materials is described.
After the preparation of starting materials refer to chapter 2.5 “Protocol: Simultaneous isolation of
bacterial DNA and viral DNA/RNA from liquid samples” to follow step 1a) or 1b) of the protocol
to continue, unless stated otherwise.

2.3.1 Serum, plasma, other cell-free body liquids

 Always mix the sample well before extraction.
 Use up to 200 µl sample and adjust to 400 µl with Resuspension Buffer R.
2.3.2 Swabs
a) Dry Swabs
 Rinse the swabs in the lowest possible volume of PBS or Resuspension Buffer R
x 400 µl for nasopharyngeal swabs
x 600 µl for oral swabs
 Squeeze the swab to the inner wall of the vial to obtain as much sample as possible.
 Use 400 µl for the extraction protocol.
 Alternative: add 400 µl Resuspension Buffer R to the Extraction Tube and rinse the
swab directly in the dissolved lysis buffer.
b) Swabs in stabilization liquid
 Use 200 µl of the stabilization solution and adjust to 400 µl with Resuspension Buffer R.
 Rinse and squeeze the swab to the inner wall of the vial before removing it.
 Caution: Some stabilization media may interfere with the lysis reaction (if you have any
questions, please refer to the FAQ or contact support).
2.3.3 Stool samples (supernatant)
a) Extraction of nucleic acids from viruses
 Transfer 100 µl / 100 mg stool sample into a 2 ml vial.
 Add 900 µl Resuspension Buffer R.
 Vortex for 30 sec, centrifuge for 1 min at 12,000 x g.
 Use up to 200 µl supernatant and adjust the volume to 400 µl with Resuspension Buffer
R.
 Avoid solid particles in the sample.
b) Extraction of bacterial DNA
 Transfer 100 µl / 100 mg stool sample into a 2 ml vial.
 Add 300 µl Resuspension Buffer R.
 Vortex for 30 sec, centrifuge for 1 min at 1,000 x g.
 Use up to 200 µl supernatant and adjust the volume to 400 µl with Resuspension Buffer
R.
 Avoid solid particles in the sample.
2.3.4 Cultivated bacteria
 Transfer 1 ml of the culture to a 2.0 ml Safe-Lock-Tube.
 Centrifuge for 2 min at 10,000 x g and completely remove the supernatant.
 Resuspend the pellet in 400 µl Resuspension Buffer R.

RTP® Pathogen Kit 11 DE 806.02_EN

 2.3.5 Urine
 Recommended volume: 15 - 50 ml depending on application and bacterial load.
 Centrifuge to sediment the bacteria and completely remove the supernatant (urea
contamination can inhibit PCR).
 Resuspend pellet in 400 µl Resuspension Buffer R.
 Alternatively, use up to 200 µl of fresh urine and adjust to 400 µl with Resuspension
Buffer R.

Note: For some applications, fresh urine can be used directly: Use up to 200 µl fresh urine,
adjust volume to 400 µl with Resuspension Buffer R and start with the extraction protocol.

2.3.6 Respiratory samples: tracheal, bronchoalveolar lavage (BAL) and

sputum
a) Non-viscous or low viscosity samples
 Mix well.
 Use up to 200 µl sample and adjust to 400 µl with Resuspension Buffer R.
b) Isolation of bacteria from viscous samples
 Sputum: transfer 150 µl + 150 µl of saturated acetylcysteine (ACC) solution
 Tracheal/BAL: transfer 1 ml + 1 ml of saturated acetylcysteine (ACC) solution.
Note: the sample/buffer ratio should be 1:1
 Incubate for 10 min at 95°C with continuous agitation.
 Centrifuge at 10,000 × g for 5 min.
 Discard supernatant, resuspend bacterial pellet in 400 µl PBS or Resuspension Buffer
R.
c) Isolation of viral DNA/RNA from viscous samples
 Transfer 150 µl of the sample into a Safe-Lock-Tube and add 150 µl saturated
acetylcysteine (ACC) (ratio sample to buffer must be 1:1).
 Incubate for 10 min at 95°C while continuously shaking.
 Allow the sample to cool down.
 Use up to 200 µl sample and adjust to 400 µl with Resuspension Buffer R.
2.3.7 Tissue Biopsies
 Add 400 µl Resuspension Buffer R to the Extraction Tube.
 Add 1-10 mg tissue biopsy sample to the dissolved lysis buffer.
 For tissues that are difficult to lyse (cartilage, kidney, or heart muscle), disruption
by bead beating with Zirconia beads is recommended.
 After mechanical treatment, incubate for 10 min at 65°C while continuously shaking.
 Centrifuge for 1 min at 10,000 x g, transfer supernatant to a new tube.
 Avoid solid particles.
 Continue with step 2 of the protocol.

2.3.8 Cell culture supernatants

 Use up to 200 µl sample and adjust to 400 µl with Resuspension Buffer R.

RTP® Pathogen Kit 12 DE 806.02_EN

 2.4 Short protocol RTP® Pathogen Kit
Refer to chapter 2.3 “Preparation of starting material” for sample specific pre-treatment.
Lyse samples
1. Add 400 µl of volume adjusted sample material to the Extraction Tube
Perform the following heating steps on a thermomixer while
continuously shaking:

a) Simultaneous isolation of bacterial and viral nucleic acids:

Incubate for 10 min at 37°C.
Incubate for 15 min at 65°C.
Optional: Incubate 10 min at 95°C.

b) Isolation of viral nucleic acids:

Incubate for 10 min at 65°C.
Optional: Incubate for 10 min at 95°C.
Optional: Add the extraction control after lysis.

Bind nucleic acids

2. Add 400 µl Binding Solution, mix by pipetting up and down or vortexing.
Transfer the sample into the RTA Spin Filter Set.
Incubate for 1 min at room temperature.
Centrifuge for 2 min at 11,000 x g.

Discard the RTA Receiver Tube with filtrate and place the RTA Spin Filter
into a new RTA Receiver Tube.

Wash to remove residual contamination

3. Add 500 µl Wash Buffer R1, centrifuge for 1 min at 11,000 x g.
Discard the RTA Receiver Tube with the filtrate and place the RTA Spin
Filter into a new RTA Receiver Tube.

4. Add 700 µl Wash Buffer R2, centrifuge 1 min at 11,000 x g.

Discard the RTA Receiver Tube with the filtrate and place the RTA Spin
Filter into a new RTA Receiver Tube.

5. Centrifuge for 4 min at maximum speed to remove residual

ethanol. Discard the RTA Receiver Tube with filtrate.

Elute nucleic acids

6. Place the Spin Filter into a 1.5 ml Receiver Tube.
Add 60 µl Elution Buffer R (preheated to 65°C) directly onto the RTA Spin
Filter.
Incubate 1 min at RT and centrifuge 1 min at 11,000 x g.
Discard the RTA Spin Filter and store eluted nucleic acids on ice.

RTP® Pathogen Kit 13 DE 806.02_EN

 2.5 Protocol for simultaneous isolation of bacterial DNA and viral
DNA/RNA from liquid samples
Please refer to chapter 2.3 “Preparation of starting material” for sample specific pre-
treatment.

1. Add 400 µl sample material to the Extraction Tube. Depending on the starting material
the sample volume must be adjusted to 400 µl with Resuspension Buffer R or PBS
buffer, vortex shortly.
Depending on sample type and target nucleic acid, perform steps in a) or b) on a thermo
shaker while continuously shaking:
a) Isolation of bacterial DNA or simultaneous isolation of bacterial and viral nucleic
acids
Incubate 10 min at 37°C. Incubate 15 min at 65°C.
For difficult to lyse bacteria (e.g., Mycobacteria) or tissue additional incubation for
10 min. at 95°C is recommended.
b) Isolation of viral nucleic acids
Incubate 10 min at 65°C.
For difficult to lyse samples like tissue an additional incubation for 10 min at 95°C is
recommended.
Note: If you want to add nucleic acids for extraction control, please add them now,
before the binding step.

2. Add 400 µl Binding Solution and mix the sample completely by pipetting up and down or
by vortexing.
Transfer the sample into the RTA Spin Filter Set and incubate for 1 min. Centrifuge for 2
min at 11,000 x g.
Discard the RTA Receiver Tube with filtrate and place the RTA Spin Filter into a new
RTA Receiver Tube.
3. Add 500 µl Wash Buffer R1 and centrifuge 1 min at 11,000 x g.
Discard the RTA Receiver Tube with filtrate and place the RTA Spin Filter into a new
RTA Receiver Tube.
4. Add 700 µl Wash Buffer R2 to the RTA Spin Filter and centrifuge 1 min at 11,000 x g.
Discard the RTA Receiver Tube with filtrate and place the RTA Spin Filter into a new
RTA Receiver Tube.
5. Remove the residual ethanol by final centrifugation for 4 min at maximum speed. Discard
the RTA Receiver Tube with filtrate.
6. Place the Spin Filter into a 1.5 ml Receiver Tube and add 60 µl of the Elution Buffer R
(preheated to 65°C) directly onto the RTA Spin Filter surface.
Incubate 1 min at RT and centrifuge 1 min at 11,000 x g.

RTP® Pathogen Kit 14 DE 806.02_EN

 3. Warranty
Invitek Diagnostics guarantees the correct functioning of the kit for the applications described in
this manual and in accordance with its intended use. In accordance with Invitek Diagnostics'
Quality Management System, certified to EN ISO 13485, the performance of all kit components
has been tested to ensure product quality. Any problems, incidents or defects should be reported
to Invitek Diagnostics immediately upon detection. Upon receipt, inspect the product to ensure
it is complete and undamaged. In case of discrepancies, the user must immediately inform
Invitek Diagnostics in writing. Modifications to the kit and protocols, as well as uses that deviate
from the original purpose, are not covered by any warranty. In case of serious incidents, the
user must also inform the competent authorities, as required by applicable legislation. Invitek
Diagnostics reserves the right to change, alter or modify any product to improve its performance
and design at any time. Invitek Diagnostics guarantees the products as set out in the General
Terms and Conditions available at [Link]. If you have any questions, please contact
techsupport@[Link].

4. Safety instructions
Ensure that anyone using this product has received instruction in general laboratory safety
practices as well as the safety information provided in this document.

x Always wear protective clothing, disposable gloves, and safety goggles when
handling chemicals.
x Replace pipette tips after each liquid transfer, preferably with an aerosol barrier, to
prevent cross-contamination.
x Do not reuse consumables.
x Discard contaminated gloves.

x Do not combine components from different kits.

x Avoid microbial contamination of reagents.
x Handle samples in a laminar flow chamber until lysis to reduce the risk of infection
from potentially infectious materials.

Before handling chemicals, read and understand all applicable safety data sheets (SDS).
These are available at [Link].
Dispose of kit waste and residual fluids in accordance with your country's regulations and
refer back to the SDS. Invitek Diagnostics has not tested the liquid waste generated by the
kit for residual infectious materials. Although contamination of liquid waste with residual
infectious materials is highly unlikely, it cannot be completely ruled out. Therefore, liquid
waste should be considered infectious and should be handled and disposed of in accordance
with applicable safety regulations.

The European Community risk and safety phrases described below refer exclusively to the
components of the RTP® Pathogen Kit that present identified risks. All safety information is
described in detail in the safety data sheet (SDS) and product instructions. The qualitative
and quantitative composition of the reagents, as well as their respective risks, is presented
in the table below.

RTP® Pathogen Kit 15 DE 806.02_EN

 4.1 Reagent Specifications and Hazard Classification
Pictogram and Hazard Hazard Precautionary
Kit Components Main Composition
Class GSH1 warnings1 Statements

50% - 100% Ammonium Chloride P264; P280;

H302; H315; P305+P351+P338+P310;
10% - 20% Hexadecyltrimethylammonium
Extraction Tubes GHS05 GHS07 GHS08 H318; H373; P314; P321; P391
bromide
GHS09 H410
0.1% - 1% Proteinase K EUH208*
Hazard
Resuspension Buffer R 100% ultrapure water

P260; P264; P270; P271;

P273; P280;
H302+H332;
Wash Buffer R1 45% - 55% Guanidinium thiocyanate P301+P330+P331;
GHS05... GHS07 H314; H412.
P303+P361+P353;
Hazard P304+P340; P501.

0.1% - 1% Sodium chloride

Wash Buffer R2 0.1% - 1% Tris HCl
0.01% - 0.1% Tris Base
Elution Buffer R 100% Ultrapure water
1
Classification and labelling according to Regulation (EC) No 1272/2008 [CLP]
GHS: Globally Harmonised System of Classification and Labelling of Chemicals
*Other EU supplementary hazard statements that do not have direct equivalents in GHS and have therefore been retained in the European Union's CLP Regulation.

Emergency medical information can be obtained 24 hours a day from infotrac, [Link]:
outside the USA: 1-352-323-3500
in the USA: 1-800-535-5053

RTP® Pathogen Kit 16 DE 806.02_EN

 4.2 List of Hazard Warnings and Precautionary Statements
Codes Hazard warning/Precautionary statement
H302 Harmful if swallowed
H314 Causes severe skin burns and eye damage.
H315 Causes skin irritation
H318 Causes serious eye damage.
H373 May cause damage to organs through prolonged or repeated exposure
H410 Very toxic to aquatic life with long-lasting effects
H412 Harmful to aquatic organisms with long-lasting effects.
P260 Do not breathe dust/fumes/gas/mist/vapours/spray.
P264 Wash hands, forearms and face thoroughly after handling.
P270 Do not eat, drink or smoke when using this product.
P273 Avoid release to the environment.
Wear protective gloves/protective clothing/eye protection/face
P280
protection/hearing protection.
IF SWALLOWED: If you feel unwell, contact a POISON CONTROL
P301+P312
CENTRE or doctor.
P301+P330+P331 IF SWALLOWED: Rinse mouth. Do NOT induce vomiting.
IF ON SKIN (or hair): Remove all contaminated clothing immediately.
P303+P361+P353
Rinse skin with water.
IF INHALED: Remove person to fresh air and keep at rest in a position
P304+P340
comfortable for breathing.
IF IN EYES: Rinse cautiously with water for several minutes. Remove
P305+P351+P338+P310 contact lenses, if present and easy to do. Continue rinsing. Immediately
call a POISON CENTRE or doctor/physician.
P314 If you feel unwell, seek medical advice.
P321 Specific treatment (see additional first aid instructions on this label).
P391 Collect spillage.
Dispose of contents/container to a hazardous or special waste collection
P501 point in accordance with local, regional, national and/or international
regulations.
Contains Proteinase, Tritirachium album serine (39450-01-6). May cause
EUH208
an allergic reaction.

RTP® Pathogen Kit 17 DE 806.02_EN

 5. Appendix
5.1 Troubleshooting

Problem Possible cause Recommendation

Low amount Insufficient cell lysis Increase lysis time.

of nucleic Continuous shaking improves lysis efficiency.
acids Reduce amount of starting material to avoid column
overload.

Incomplete elution Increase incubation time with preheated Elution Buffer R

to 5-10 min.
Elute twice with 100 µl Elution Buffer R.
Use higher volume of Elution Buffer R.

Low nucleic acid- Elute nucleic acids with a lower volume of Elution
concentration in the Buffer R, do not use volumes below 40 µl.
sample

Incorrect storage of starting Ensure that starting material is appropriately stored.

material Avoid repeated thaw-freeze cycles of the sample material.

Wash Buffers were Ensure, that the correct amount of ethanol/isopropanol is

incorrectly prepared added to the Wash Buffers and that all solutions are
stored firmly closed.

Degraded Ensure that the starting material is stored at appropriate

nucleic acids Old material conditions (–20°C/-80°C).

Ethanol carryover during Increase time of drying step for removal of ethanol.
elution

Nucleic acids Salt carry-over during Check the Wash Buffers for salt precipitates. If there are
do not elution any precipitates visible, solve them by carefully warming
perform well up to 30°C.
in Ensure that the Wash Buffers are at room temperature
downstream before use.
applications
(e.g. real-time
PCR or NGS) Insufficient cell lysis See above.

Coloured Inefficient washing Repeat washing step.

residues on
the RTA Spin
filter after Wash Buffers were
incorrectly prepared See above.
washing

RTP® Pathogen Kit 18 DE 806.02_EN

 5.2 Other documents and further information
Visit [Link] for further information on:

x FAQs and troubleshooting tips

x Manuals in different languages
x Safety data Sheets (MSDS)
x Web support
x Product videos

If, despite careful study of the operating instructions and further information, you still
require assistance, please contact us at techsupport@[Link] or the distributor
responsible for you.

5.3 Ordering information

Product Package Size Catalogue number
RTP® Pathogen Kit 50 preparations 1040500200
RTP® Pathogen Kit 250 preparations 1040500300

Revision history

Revision Date Description

DE 806.01_EN 2025-05-27 New document

DE 806.02_EN 2025-09-15 New structure and addition of

performance characteristics

RTP® Pathogen Kit 19 DE 806.02_EN

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