Romed Coronavirus Antigen Rapid Test Cassette Instruction Manual
- June 5, 2024
- Romed
Table of Contents
- Romed Coronavirus Antigen Rapid Test Cassette
- INTENDED USE
- SUMMARY AND EXPLANATION
- PRINCIPLE OF THE TEST
- WARNINGS AND PRECAUTIONS
- STORAGE AND STABILITY
- SPECIMEN COLLECTION
- SAMPLE PREPARATION PROCEDURE
- SPECIMEN TRANSPORT AND STORAGE
- TEST PROCEDURE
- QUALITY CONTROL
- PERFORMANCE CHARACTERISTICS
- Limit of Detection (LOD)
- Microbial Interference
- INDEX OF SYMBOLS
- Documents / Resources
Romed Coronavirus Antigen Rapid Test Cassette
INTENDED USE
The Coronavirus Ag Rapid Test Cassette (Swab) is an in vitro immunochromatographic assay for the qualitative detection of nucleocapsid protein antigen from SARS-CoV-2 in direct nasopharyngeal (NP) swab or nasal swab specimens directly from individuals who are suspected of COVID-19 by their healthcare provider within the first ten days of symptom onset, and asymptomatic individuals. It is intended to aid in the rapid diagnosis of SARS-CoV-2 infections. Negative results from patients with symptom onset beyond ten days should be treated as presumptive and confirmation with a molecular assay, if necessary, for patient management, may be performed. The Coronavirus Ag Rapid Test Cassette (Swab) does not differentiate between SARS- CoV and SARS-CoV-2. Coronavirus Ag Rapid Test Cassette (Swab) is intended for use by healthcare professionals or trained operators who are proficient in performing rapid tests and trained clinical laboratory personnel specifically instructed on in vitro diagnostic procedures and proper infection control procedures or individuals similarly trained in point of care settings.
SUMMARY AND EXPLANATION
The novel coronaviruses belong to the β genus. COVID-19 is an acute
respiratory infectious disease. People are generally susceptible. Currently,
the patients infected by the novel coronavirus are the main source of
infection; asymptomatic infected people can also be an infectious source.
Based on the current epidemiological investigation, the incubation period is 1
to 14 days, mostly 3 to 7 days. The main manifestations include fever,
fatigue, and dry cough. Nasal congestion, runny nose, sore throat, myalgia,
and diarrhea are found in a few cases.
This test is for detection of SARS-CoV-2 nucleocapsid protein antigen. Antigen
is generally detectable in upper respiratory specimens during the acute phase
of infection. Rapid diagnosis of SARS-CoV-2 infection will help healthcare
professionals to treat patients and control the disease more efficiently and
effectively.
To effectively monitor the SARS-CoV-2 pandemic, systematic screening and
detection of both clinical and asymptomatic COVID-19 cases is critical.
Particularly, the identification of subclinical or asymptomatic cases is
important to reduce or stop the infection because these individuals may
transmit the virus. Coronavirus Ag Rapid Test Cassette (Swab) allows effective
screening of COVID-19 infection.
PRINCIPLE OF THE TEST
The Coronavirus Ag Rapid Test Cassette (Swab) is an immunochromatographic membrane assay that uses highly sensitive monoclonal antibodies to detect nucleocapsid protein from SARS-CoV-2 in direct nasopharyngeal (NP) swab or nasal swab. The test strip is composed of the following parts: namely sample pad, reagent pad, reaction membrane, and absorbing pad. The reagent pad contains the colloidal-gold conjugated with the monoclonal antibodies against the nucleocapsid protein of SARS-CoV-2; the reaction membrane contains the secondary antibodies for nucleocapsid protein of SARS-CoV-2. The whole strip is fixed inside a plastic device. When the sample is added into the sample well, conjugates dried in the reagent pad are dissolved and migrate along with the sample. If SARS-CoV-2 nucleocapsid antigen is present in the sample, a complex forms between the anti-SARS-2 conjugate and the virus will be captured by the specific anti-SARS-2 monoclonal antibodies coated on the test line region (T). Absence of the test line (T) suggests a negative result. To serve as a procedural control, a red line will always appear in the control line region (C) indicating that proper volume of sample has been added and membrane wicking has occurred.
MATERIALS PROVIDED
- 20 Test Cassettes
- 2 Extraction Buffer Vials
- 20 Sterile Swabs
- 20 Extraction Tubes and Tips
- 1 Workstation
- 1 Package Insert
MATERIALS REQUIRED BUT NOT PROVIDED
Clock, timer, or stopwatch
WARNINGS AND PRECAUTIONS
- For in vitro diagnostic use only.
- The test device should remain in the sealed pouch until use.
- Do not use kit past its expiration date.
- Swabs, tubes, and test devices are for single use only.
- Solutions that contain sodium azide may react explosively with lead or copper plumbing. Use large quantities of water to flush discarded solutions down a sink.
- Do not interchange or mix components from different kit lots.
- Testing should only be performed using the swabs provided within the kit.
- To obtain accurate results, do not use visually bloody or overly viscous samples.
- If the test is carried out by or being supervised by a healthcare professional or trained individual, it is recommended they wear appropriate PPE, whist changing gloves between patients. The patient themselves does not need to wear PPE.
- Specimens must be processed as indicated in the SPECIMEN COLLECTION and SAMPLE PREPARATION PROCEDURE sections of this Product Insert. Failure to follow the instructions for use can result in inaccurate results.
- Proper laboratory safety techniques should be followed at all times when working with SARS-CoV-2 patient samples. Patient swabs used Test Strips and used extraction buffer vials may be
- potentially infectious. Proper handling and disposal methods should be established by the laboratory in accordance with local regulatory requirements.
- Inadequate or inappropriate specimen collection and storage can adversely affect results.
- Humidity and temperature can adversely affect results.
- Dispose of test device and materials as biohazardous waste in accordance with federal, state, and local requirements.
STORAGE AND STABILITY
- The kit can be stored at room temperature or refrigerated (2-30°C).
- Do not freeze any of the test kit components.
- Do not use test device and reagents after expiration date.
- Test devices that have been outside of the sealed pouch for more than 1 hour should be discarded.
- Close the kit box and secure its contents when not in use.
SPECIMEN COLLECTION
Nasopharyngeal Swab
- Using the sterile swab provided in the kit, carefully insert the swab in the patient’s nostril.
- Swab over the surface of the posterior nasopharynx and rotate the swab several times.
- Withdraw the swab from the nasal cavity. The specimen is now ready for preparation using the extraction buffer provided in the test kit.
Nasal Swab
- Using the sterile swab provided in the kit, carefully insert the swab into one nostril of the patient. The swab tip should be inserted up to 2-4 cm until resistance is met.
- Roll the swab 5 times along the mucosa inside the nostril to ensure that both mucus and cells are collected.
- Using the same swab, repeat this process for the other nostril to ensure that an adequate sample is collected from both nasal cavities.
- Withdraw the swab from the nasal cavity. The specimen is now ready for preparation using the extraction buffer provided in the test kit.
SAMPLE PREPARATION PROCEDURE
- Insert the test extraction tube into the workstation in the kit. Make sure that the tube is standing firm and reaches the bottom of the workstation.
- Add 0.3 ml (about 10 drops) of the sample extraction buffer into the extraction tube.
- Insert the swab into the extraction tube which contains 0.3 ml of the extraction buffer.
- Roll the swab at least 6 times while pressing the head against the bottom and side of the extraction tube.
- Leave the swab in the extraction tube for 1 minute
- Squeeze the tube several times with fingers from outside of the tube to immerse the swab. Remove the swab.
- Fit the dropper tip with filter on top of the extraction tube tightly.
SPECIMEN TRANSPORT AND STORAGE
Do not return the sterile swab to the original paper packaging.
Specimen should be tested immediately after collection. If immediate testing
of specimen is not possible, insert the swab into an unused general-purpose
plastic tube. Ensure the breakpoint swab is level with the tube opening. Bend
the swab shaft at a 180 degrees angle to break it off at the breaking point.
You may need to gently rotate the swab shaft to complete the breakage. Ensure
the swab fits within the plastic tube and secure a tight seal. The specimen
should be disposed and recollected for retesting if untested for longer than 1
hour.
TEST PROCEDURE
Allow the test device, test sample and buffer to equilibrate to room temperature (15-30°C) prior to testing.
- Just prior to testing remove the test device from the sealed pouch and lay it on a flat surface.
- Push the nozzle which contains the filter onto the extraction tube. Ensure the nozzle has a tight fit.
- Hold the extraction tube vertically and add 4 drops (approximately 100 μL) of test sample solution into the sample well.
- Start the timer.
- Read the results at 15 minutes. Do not interpret the result after 20 minutes.
INTERPRETATION OF RESULTS
-
POSITIVE:
The presence of two lines as control line (C) and test line (T) within the result window indicates a positive result. -
NEGATIVE:
The presence of only control line (C) within the result window indicates a negative result. -
INVALID:
If the control line (C) is not visible within the result window after performing the test, the result is considered invalid. Some causes of invalid results are because of not following the directions correctly or the test may have deteriorated beyond the expiration date. It is recommended that the specimen be re-tested using a new test
NOTE:
- The intensity of colour in the test line region (T) may vary depending on the concentration of analyses present in the specimen. Therefore, any shade of colour in the test line region (T) should be considered positive. This is a qualitative test only and cannot determine the concentration of analytes in the specimen.
- Insufficient specimen volume, incorrect operating procedure or expired tests are the most likely reasons for control band failure.
QUALITY CONTROL
A procedural control is included in the test. A red line appearing in the control line region (C) is the internal procedural control. It confirms sufficient specimen volume and correct procedural technique. Control standards are not supplied with this test. However, it is recommended that positive and negative controls are sourced from a local competent authority and tested as a good laboratory practice, to confirm the test procedure and verify the test performance.
LIMITATIONS
- The etiology of respiratory infection caused by microorganisms other than SARS-CoV-2 will not be established with this test. The Coronavirus Ag Rapid Test Cassette (Swab) can detect both viable and non-viable SARS-CoV-2 The performance of the Coronavirus Ag Rapid Test Cassette (Swab) depends on antigen load and may not correlate with viral culture results performed on the same specimen.
- Failure to follow the Test Procedure may adversely affect test performance and/or invalidate the test result.
- If the test result is negative and clinical symptoms persist, additional testing using other clinical methods is recommended. A negative result does not at any time rule out the presence of SARS-CoV-2 antigens in specimen, as they may be present below the minimum detection level of the test or if the sample was collected or transported improperly.
- As with all diagnostic tests, a confirmed diagnosis should only be made by a physician after all clinical and laboratory findings have been evaluated.
- Positive test results do not rule out co-infections with other pathogens.
- Positive test results do not differentiate between SARS-CoV and SARS-CoV-2.
- The amount of antigen in a sample may decrease as the duration of illness increases. Specimens collected after day 10 of illness are more likely to be negative compared to a RT-PCR assay.
- Negative results from patients with symptom onset beyond ten days, should be treated as presumptive and confirmation with a molecular assay, if necessary, for patient management, may be performed.
- Negative results do not rule out SARS-CoV-2 infection and should not be used as the sole basis for treatment or patient management decisions, including infection control decisions.
PERFORMANCE CHARACTERISTICS
Clinical Sensitivity, Specificity and Accuracy
Nasopharyngeal Swab
Clinical Performance of the Coronavirus Ag Rapid Test Cassette (Swab) was
evaluated by being involved in 7 sites within the US where patients were
enrolled and tested. Testing was performed by 24 Healthcare Workers that were
not familiar with the testing procedure. A total of 865 fresh nasopharyngeal
swab samples was collected and tested, which includes 119 positive samples and
746 negative samples. The Coronavirus Ag Rapid Test Cassette (Swab) results
were compared to USFDA Emergency Use Authorized RT-PCR assays for SARS-CoV-2
in nasopharyngeal swab specimens. Overall study results are shown in Table 1.
| Method | PCR | Total Results |
|---|---|---|
| Coronavirus Ag Rapid Test Cassette | Results | Positive |
| Positive | 117 | 3 |
| Negative | 2 | 743 |
| Total Results | 119 | 746 |
Relative Sensitivity: 98.32% (95% Cl: 94.06% to 99.80%)
Relative Specificity: 99.60% (95% CI: 98.83.03% to 99.92%)
Accuracy: 99.42% (95%CI*: 98.66% to 99.81%)
Nasal Swab
A total of 237 fresh nasal swab samples was collected and tested, which
includes 109
positive samples and 128 negative samples. The Coronavirus Ag Rapid Test
Cassette (Swab) results were compared to results of USFDA Emergency Use
Authorized RT-PCR
assays for SARS-CoV-2 in Nasopharyngeal swab specimens. Overall study results
are shown in Table 2.
Table 2: Coronavirus Ag Rapid Test Cassette (Swab) (Nasal Swab) vs PCR
| Method | PCR | Total Results |
|---|---|---|
| Coronavirus Ag Rapid Test Cassette | Results | Positive |
| Positive | 106 | 0 |
| Negative | 3 | 128 |
| Total Results | 109 | 128 |
Relative Sensitivity: 97.25% (95% Cl: 92.17% to 99.43%)
Relative Specificity: 100% (95% CI: 97.16% to 100%)
Accuracy: 98.73% (95%CI: 96.35% to 99.74%)
Confidence Intervals
Limit of Detection (LOD)
LOD studies determine the lowest detectable concentration of SARS-CoV-2 at which approximately 95% of all (true positive) replicates test positive. Heat inactivated SARS- CoV-2 virus, with a stock concentration of 4.6×105 TCID50 / mL, was spiked into negative specimen and serially diluted. Each dilution was ran in triplicate on the Coronavirus Ag test. The Limit of Detection of the Coronavirus Ag Rapid Test Cassette (Swab) is 1.15×102 TCID50 / mL (Table 3).
Table 3: Limit of Detection (LOD) Study Results
| Concentration | No. Positive/Total | Positive Agreement |
|---|---|---|
| 1.15 x 102 TCID50 / mL | 180/180 | 100% |
High Dose Hook Effect
No high dose hook effect was observed when testing up to a concentration of
4.6 x 105 TCID50 / mL of heat inactivated SARS-CoV-2 virus.
Cross Reactivity
Cross reactivity with the following organisms has been studied. Samples
positive for the following organisms were found negative when tested with the
Coronavirus Ag Rapid Test Cassette (Swab).
| Pathogens | Concentration |
|---|---|
| Respiratory syncytial virus Type A | 5.5×107 PFU/mL |
| Respiratory syncytial virus Type B | 2.8×105 TCID50/mL |
| Novel influenza A H1N1 virus (2009) | 1×106 PFU/mL |
| Seasonal influenza A H1N1 virus | 1×105 PFU/mL |
| Influenza A H3N2 virus | 1×106 PFU/mL |
| Influenza A H5N1 virus | 1×106 PFU/mL |
| Influenza B Yamagata | 1×105 PFU/mL |
| Influenza B Victoria | 1×106 PFU/mL |
| Rhinovirus | 1×106 PFU/mL |
| Adenovirus 3 | 5×107.5 TCID50/mL |
| Adenovirus 7 | 2.8×106 TCID50/mL |
| EV-A71 | 1×105 PFU/mL |
| Mycobacterium tuberculosis | 1×103 bacteria/mL |
| Mumps virus | 1×105 PFU/mL |
| Human coronavirus 229E | 1×105 PFU/mL |
| Human coronavirus OC43 | 1×105 PFU/mL |
| Human coronavirus NL63 | 1×106 PFU/mL |
| Human coronavirus HKU1 | 1×106 PFU/mL |
| Parainfluenza virus 1 | 7.3×106 PFU/mL |
| Parainfluenza virus 2 | 1×106 PFU/mL |
| --- | --- |
| Parainfluenza virus 3 | 5.8×106 PFU/mL |
| Parainfluenza virus 4 | 2.6×106 PFU/mL |
| Haemophilus influenzae | 5.2×106 CFU/mL |
| Streptococcus pyogenes | 3.6×106 CFU/mL |
| Streptococcus pneumoniae | 4.2×106 CFU/mL |
| Candida albicans | 1×107 CFU/mL |
| Bordetella pertussis | 1×104 bacteria/mL |
| Mycoplasma pneumoniae | 1.2×106 CFU/mL |
| Chlamydia pneumoniae | 2.3×106 IFU/mL |
| Legionella pneumophila | 1×104 bacteria/mL |
| Staphylococcus aureus | 3.2×108 IFU/mL |
| Staphylococcus epidermidis | 2.1×108 IFU/mL |
Interfering Substance
The following substances, naturally present in respiratory specimens or that
may be artificially introduced into the nasal cavity or nasopharynx, were
evaluated with the Coronavirus Ag Rapid Test Cassette (Swab) at the
concentrations listed below and were found not to affect test performance.
| Substance | Concentration |
|---|---|
| Human blood (EDTA anticoagulated) | 20% (v/v) |
| Mucin | 5 mg/mL |
| Oseltamivir phosphate | 5 mg/mL |
| Ribavirin | 5 mg/mL |
| Levofloxacin | 5 mg/mL |
| Azithromycin | 5 mg/mL |
| Meropenem | 5 mg/mL |
| Tobramycin | 2 mg/mL |
| Phenylephrine | 20% (v/v) |
| Oxymetazoline | 20% (v/v) |
| 0.9% sodium chloride | 20% (v/v) |
| A natural soothing ALKALOL | 20% (v/v) |
| Beclomethasone | 20% (v/v) |
| Hexadecadrol | 20% (v/v) |
| Flunisolide | 20% (v/v) |
| Triamcinolone | 20% (v/v) |
| Budesonide | 20% (v/v) |
| Mometasone | 20% (v/v) |
| Fluticasone | 20% (v/v) |
| Fluticasone propionate | 20% (v/v) |
Microbial Interference
To evaluate whether potential microorganisms in clinical samples interfere with the detection of Coronavirus Ag Rapid Test Cassette (Swab) so as to produce false negative results. Each pathogenic microorganism was tested in triplicate in the presence of heat inactivated SARS-Cov-2 virus (2.3×102 TCID50 / mL). No cross reactivity or interference was seen with the microorganisms listed in the table below.
| Pathogens | Concentration |
|---|---|
| Respiratory syncytial virus Type A | 5.5×107 PFU/mL |
| Respiratory syncytial virus Type B | 2.8×105 TCID50/mL |
| Novel influenza A H1N1 virus (2009) | 1×106 PFU/mL |
| Seasonal influenza A H1N1 virus | 1×105 PFU/mL |
| Influenza A H3N2 virus | 1×106 PFU/mL |
| Influenza A H5N1 virus | 1×106 PFU/mL |
| Influenza B Yamagata | 1×105 PFU/mL |
| Influenza B Victoria | 1×106 PFU/mL |
| Rhinovirus | 1×106 PFU/mL |
| Adenovirus 1 | 1×106 PFU/mL |
| Adenovirus 2 | 1×105 PFU/mL |
| Adenovirus 3 | 5×107.5 TCID50/mL |
| Adenovirus 4 | 1×106 PFU/mL |
| Adenovirus 5 | 1×106 PFU/mL |
| Adenovirus 7 | 2.8×106 TCID50/mL |
| EV-A71 | 1×105 PFU/mL |
| EV-B69 | 1×105 PFU/mL |
| EV-C95 | 1×105 PFU/mL |
| EV-D70 | 1×105 PFU/mL |
| Mycobacterium tuberculosis | 1×103 bacteria/mL |
| Mumps virus | 1×105 PFU/mL |
| Human coronavirus 229E | 1×105 PFU/mL |
| Human coronavirus OC43 | 1×105 PFU/mL |
| Human coronavirus NL63 | 1×106 PFU/mL |
| Human coronavirus HKU1 | 1×106 PFU/mL |
| Human Metapneumovirus (hMPV) | 1×106 PFU/mL |
| Parainfluenza virus 1 | 7.3×106 PFU/mL |
| Parainfluenza virus 2 | 1×106 PFU/mL |
| Parainfluenza virus 3 | 5.8×106 PFU/mL |
| Parainfluenza virus 4 | 2.6×106 PFU/mL |
| Haemophilus influenzae | 5.2×106 CFU/mL |
| Streptococcus pyogenes | 3.6×106 CFU/mL |
| Streptococcus agalactiae | 7.9×107 CFU/mL |
| Streptococcus pneumoniae | 4.2×106 CFU/mL |
| Pooled human nasal wash | N/A |
| Candida albicans | 1×107 CFU/mL |
| --- | --- |
| Bordetella pertussis | 1×104 bacteria/mL |
| Mycoplasma pneumoniae | 1.2×106 CFU/mL |
| Chlamydia pneumoniae | 2.3×106 IFU/mL |
| Legionella pneumophila | 1×104 bacteria/mL |
INDEX OF SYMBOLS
VAN OOSTVEEN MEDICAL B.V. ROMED – HOLLAND HERENWEG 269
3648 CH WILNIS
THE NETHERLANDS
WWW.ROMED.NL
V006, 2021-02
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