ProGnosis Lateral Flow Test for COVID-19 / Ag 2019-nCoV Instructions For Use
- June 2, 2024
- ProGnosis Biotech
Table of Contents
RAPID TEST Ag 2019-nCoV
This Lateral Flow test kit is manufactured by ProGnosis Biotech S.A. and complies with the specifications on the Standard EN ISO 13485:2016
Use only the current version of Product Data Sheet enclosed with the kit.
Rapid Test Ag 2019-nCoV, V1310/V1330, is a qualitative Lateral Flow test for
the detection of SARS-CoV-2 antigen in nasopharyngeal specimen.
The Lateral flow kit contains all reagents required for the immunoassay
method.
Specimen:
Nasopharyngeal swab.
- For in vitro diagnostic use only
- Rapid Test Ag 2019-nCoV is a sensitive screening assay for the detection of SARSCoV-2. Results should not be used as the only source to diagnose or to determine infection status.
- Negative results do not rule out SARS-CoV-2 infection
- Test should only be conducted by medical personnel
- Test time (incubation time after specimens preparation): 15 min
- Shelf life: 12 months
- Storage: 4-30oC
1. Description
Rapid Test Ag 2019-nCov is a qualitative, lateral flow immunoassay designed to
detect the presence or absence of nucleocapsid protein of SARS-CoV-2 in
nasopharyngeal swab specimens directly collected.
The antigen (Nucleocapsid Protein, NP) is generally detectable during the
acute phase of infection.
2. Principal of the assay
In this test, antibodies specific to the NP are coated on the test line region of the nitrocellulose membrane. During testing, antigens of SARS-CoV-2 in the specimen react with the antibodies that are coated onto gold nanoparticles. The mixture migrates up the membrane to react with the antibodies immobilized on the membrane and generate one colored line in the test region. The presence of this colored line indicates a positive result. To serve as a procedural control, a colored line will always appear in the control region if the test has been performed properly.
3. Introduction
A novel coronavirus (identified as 2019-nCoV) emerged in the Chinese province of Hubei (Wuhan) in December 2019, which has resulted in hundreds of thousands of confirmed human infections worldwide. Cases of severe illness and deaths have been reported. On February 11, 2020 the International Committee for Taxonomy of Viruses (ICTV) renamed the virus SARS-CoV-2. The median incubation time is estimated to be approximately 5 days with symptoms estimated to be present within 12 days of infection. The most common symptoms of COVID-19 (according to WHO), are similar to other viral respiratory diseases and include fever, dry cough and tiredness. The virus spreads primarily through droplets of saliva or discharge from the nose when an infected person coughs or sneezes. Coronaviruses are enveloped, positive-sense, single-stranded RNA viruses with a nucleocapsid of helical symmetry and are composed of several proteins including the Spike (S), Envelope (E), Membrane (M) and Nucleocapsid (N) proteins (Figure 1). Molecular and antigen testing are the only techniques capable of detecting the SARS-CoV-2 virus. Nucleocapsid protein is a most abundant protein of coronavirus. During virion assembly, N protein binds to viral RNA and leads to formation of the helical nucleocapsid. Nucleocapsid protein is a highly immunogenic phosphoprotein also implicated in viral genome replication and in modulating cell signaling pathways. Because of the conservation of N protein sequence and its strong immunogenicity, the N protein of coronavirus is chosen as a diagnostic tool.
4. Reagents Provided
5. Materials required but not provided
- Clock or Timer
- Gloves and face protection shield
- Container for biohazardous waste
6. Storage Instructions
Store kit components between 4 and 30°C (39.2 – 96°F). Do not freeze any components provided. Reseal the unused sticks in the storage vial together with the desiccant bag provided. Expiry of the kit and reagents is stated on their labels and no quality guarantee is accepted after the expiration date. The expiry of the kit components can only be guaranteed if the components are stored properly and the reagent is not contaminated prior handling.
7. Safety and Precautions for use
7.1 Health and safety precautions
- Use gloves, protective clothing and eye/face protection and handle appropriately with the requisite Good Laboratory Practices. The product must only be used by qualified personnel.
- Dispose of all specimens and materials used to perform the test as though they contain an infectious agent. Laboratory, chemical or biohazardous wastes must be handled and discarded in accordance with the local, regional and national regulations.
- During the specimen collection human source material spills should be also treated as potentially infectious. Spills should be immediately decontaminated, including the spill area, materials and any contaminated surfaces, with an appropriate chemical disinfectant and should be wiped away.
- Do not re-use any of the kit components as they are single-use only.
- Sterile swabs must be used only for nasopharyngeal specimen. Avoid to touch the tip of the swab.
- Do not eat, drink or smoke in the area where the specimens and the kit are stored and handled.
- All positive results should be processed following local news and regulations.
7.2 Precautions related to the procedure
- In accordance with Article 1, Paragraph 2b of European Directive 98/79/EC, the use of in-vitro diagnostic medical devices is envisaged by the manufacturer to ensure the suitability, performance, and safety of these products. Consequently, the testing procedure, information, precautions, and warnings in the instructions for use must be followed rigorously. No changes to the test procedure are permitted, nor is any use in combination with other products not approved by the manufacturer. The user is solely responsible for any such changes. The manufacturer is not responsible for false results nor incidents arising as a result of these.
- Do not use the kit if the packaging of components is damaged, if there is an expired reagent or if the desiccant bag is absent inside the vial containing the sticks.
- All reagents should be warmed in room temperature before use.
- Cover or cap all reagents when not in use.
- Do not mix and interchange different specimens.
- Do not interchange individual reagents between kits of different lot numbers
8. Method Procedure
8.1 Calculate the number of swabbing sticks and tubes needed, according
to the number of samples to collect.
8.2 Mark the extraction tubes according to the specimens you intend to
collect and add 13 drops of the Running Buffer to each one of them from the
dropper bottle.
8.3 Remove a sterile nasopharyngeal swab from the pouch and place it into
one of the patient’s nostrils. When it reaches the posterior nasopharynx
rotate 3 to 5 times over the surface and then remove it slowly while rotating
it (Figure 3).
8.4 Place the swab in the extraction tube, rotate the swab forcefully
against the side of the tube for 1min. Best results are obtained when the
specimen is vigorously extracted in the solution.
8.5 Remove the swab, squeezing the sides of the tube to extract as much
liquid as possible.
8.6 Discard the swab.
8.7 Add 3 drops into the test tube and immerse the test stick following
the direction shown by the arrows, so the uncovered area of the sticks gets
soaked.
Note: In case the test stick gets inserted in the wrong direction
(arrows pointing up) and gets wet at the top label area,
it becomes useless and has to be replaced with a new test
stick.
8.8 After 15 minutes, the test stick can be visually read and interpreted
according to the corresponding figure.
Note: The test result should not be read and interpreted
after 15 minutes.
9. Interpretation of results
Positive: Two visible colored bands appear at both Test (T) and Control
(C) line. It indicates a positive result for the SARS-CoV-2 Nucleocapsid
Protein in the specimen.
Negative: One visible colored band appears at Control line. It indicates
that the concentration of the SARS-CoV-2 NP is zero or below the detection
limit of the test.
Invalid: No colored band appears at Control line no matter whether it
appears at Test line or not.
10. Limitations
- The test procedure, precautions and interpretation of results for this test must be followed strictly when testing.
- After the extraction specimens should be tested as soon as possible. Otherwise they can be stored at room temperature 20 25°C (68-77°F) for two hours. In this case use the cap to close the extraction tube without discarding the droplet.
- The test should be used for the detection of SARS-CoV-2 antigen ONLY in nasopharyngeal swab specimens.
- Failure to follow the guidelines for proper specimen collection, test procedure and interpretation of test results may adversely affect test performance and/or produce invalid result.
- USE ONLY the sterile swabs that are provided in the kit for the specimen collection.
- During specimen collection avoid contact with bleeding areasand excess of mucus of the nasopharynx as both of them may give a false positive result due to interference with the test performance.
- Positive results indicate the presence of SARS-CoV-2 antigens but a diagnosis of an infection should only be made by a physician evaluating all clinical and laboratory findings and must be based in the correlation of the results with further clinical observations.
- A negative test result may occur if the level of extracted antigen in a specimen is below the sensitivity of the test or if a poor quality specimen is obtained.
- Positive test results do not rule out co-infection with other pathogens.
- The Rapid Test Ag will indicate the presence of SARS-CoV-2 NP in the specimen from both viable and non-viable virus.
11 Immunoassay Performance
11.1 Cross-reactivity
Ιn order to determine the cross reactivity of Rapid Test Ag, an evaluation was
performed; no cross reactivity against organism, pathogens that could cause
infections
- Adenovirus
- Astrovirus
- Alpha coronavirus 229E
- Alpha coronavirus NL63
- Beta coronavirus OC43
- Beta coronavirus HKU1
- Escherichia Coli O157
- Human Transferrin
- Influenza A virus
- Influenza B virus
- Listeria monocytogenes
- Salmonella enteritidis
- Streptococcus pneumococcal
- Streptococcus pyogenes
Rapid Test Ag 2019-nCov could have some cross reaction with SARS and very low with MERS
11.2 Limit of Detection
The lowest detectable concentration of an analyte in a method is known as LOD. In this case, we check the concentration of heat inactivated SARS-CoV-2 isolate USA-WA1/2020 in Rapid Test Ag 2019-nCov. The LOD is the level at which 95% of the replicates are characterized as positive. The results of 20 replicates of 6 dilutions with heat inactivate virus are shown at the table below.
LOD : 2.87 x 103 TCID50/mL
11.3 High Dose Hook Effect
No high dose hook effect was observed up to 1.15 x 107 TCID50/mL of
inactivated SARS-CoV-2 or with the Rapid Test Ag 2019-nCoV.
11.4 Clinical performance characteristics
For the clinical performance of the Rapid Test Ag 2019-nCoV 458 negative and
90 positive specimens were tested (confirmed with RT-PCR assay SARS-COV-2
R-GENE® Biomerieux, RNeasy Mini Kit Qiagen). The results are presented at the
table below.
12. Method Summary
Total procedure time (after specimens preparation): 15 min.
Add 13 drops of the Running Buffer into the extraction tube
Place the swab into one of the patient’s nostrils. When it reaches the posterior nasopharynx rotate 3 to 5 times over the surface and then remove it slowly while rotating it
Insert the swab into the extraction tube and rotate it forcefully
against the side of the tube for 1min
Remove the swab, squeezing the sides of the tube to extract as
much liquid as possible
Add 3 drops of the extraction to the test tube
Place the test stick into the test tube with the arrows pointing down, so the uncovered area gets soaked
Wait 15 mins)
Formation of 2 lines indicates presence of SARS-CoV-2 antigen in the
specimen
13. References
-
Centers for Disease Control and Prevention. https://www.cdc.gov/coronavirus/2019-ncov/lab/
guidelines-clinical-specimens.html -
BioRxiv (https://www.biorxiv.org/content/10.1101/2020.02.07.937862v1). Accessed March 30, 2020.
-
Wu F, et al. A new coronavirus associated with human respiratory disease in China. Nature 2020;579:265-269.
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