Instructions for Milistan Combi rapid test for the detection of COVID-19 coronavirus antigen + influenza A/B No. 1
Indications for use
MILISTAN COMBI nasal test Express test for the detection of COVID-19 virus antigen and influenza A/B is an in vitro immunological assay. The assay is intended for the direct and qualitative detection of SARS-CoV-2 viral nucleoprotein antigens and influenza A/B in nasal cavity content samples.
Self-test. A negative result does not rule out the possibility of infection. A doctor's consultation is required for a definitive diagnosis.
Description
MILISTAN COMBI nasal test The rapid test for the detection of COVID-19 virus antigen and influenza A/B detects viral antigens by visual interpretation of colored bands that appear on the two test strips of the device.
COVID-19 test:
Anti-SARS-CoV-2 antibodies are immobilized on the test area of a nitrocellulose membrane. Anti-SARS-CoV-2 antibodies conjugated to colored particles are immobilized on a conjugate pad. Nasal secretions collected by the user must be mixed with the extraction buffer, which is individually packaged in the kit.
During testing, the target antigens, if present in the nasal secretions, will be released into the extraction buffer. The sample is transported by capillary action, interacts with the reagents on the sample pad, and then the target antigens will bind to the anti-SARS-CoV-2 antibodies on the conjugate pad. Thus, the antigen-antibody complex will capture the anti-SARS-CoV-2 antibodies immobilized in the test region. Excess colored particles will be captured in the control region of the membrane.
The presence of a colored band in the test zone indicates a positive result for SARS-CoV-2 viral antigens, and its absence indicates a negative result. The colored band in the control zone is intended for the control procedure, it indicates that sufficient sample has been added and the migration of fluid along the membrane has been successful.
Influenza A/B test:
Anti-influenza A and anti-influenza B antibodies are immobilized on two separate test areas of a nitrocellulose membrane. Anti-influenza A and anti-influenza B antibodies conjugated to colored particles are immobilized on a conjugate pad.
Nasal secretions collected by the user must be mixed with the extraction buffer, which is individually packaged in the kit.
During testing, the target antigens, if present in the nasal secretions, will be released into the extraction buffer. The sample moves along the strip by capillary action, it interacts with the reagents on the sample pad, and then the target antigens bind to the antibodies on the conjugate pad. Therefore, the antigen-antibody complex will be captured by the antibodies immobilized on the two test areas. Excess colored particles will be captured in the control area of the membrane.
The presence of a colored band in the test zones indicates a positive result for influenza A and B viral antigens, and its absence indicates a negative result. The colored band in the control zone is intended for the control procedure, it indicates that sufficient specimen has been added and the fluid migration along the membrane has been successful.
COMPLETENESS. Materials provided
Nasal swab pen with test strips for COV and FLU
Extraction buffer
Instructions for use
Required materials not provided
Clock, timer, or stopwatch
Reservation
For in vitro diagnostic use only
Carefully insert the device into your nostrils.
DO NOT SWALLOW
Before starting the test, read the instructions for use carefully to obtain an accurate result. You must follow their instructions.
Do not use the kit or its components after the expiration date.
The device contains animal material and should be handled as a potential biohazard. Do not use if the package is damaged or opened.
The test devices are packaged in foil pouches to prevent moisture from entering during storage. Inspect the foil pouch before opening. Do not use the devices if the foil has holes or if the pouch is not completely sealed. Erroneous results may be obtained if reagents or kit components that have been stored improperly are used.
Failure to bring specimens and reagents to room temperature prior to testing may reduce assay sensitivity. Imprecise or improper specimen collection, storage, and transportation may result in false-negative assay results.
Avoid skin contact with the buffer.
Do not pierce the foil in the extraction buffer container before starting the test.
Storage conditions
MILISTAN COMBI nasal test Express test for COVID-19 and influenza A/B virus antigen detection, when not in use, should be stored at a temperature of 230°C.
The test components are stable until the expiration date indicated on the test package.
Method of application
TEST PROCEDURE
Before use, the test kit must be brought to room temperature (1530°C).
2. Carefully insert the sample collection pad into the nasal passage to a depth of approximately 1 – 2 cm, twist 5 (five) times and remove.
Perform a similar manipulation with the second nasal passage to collect a sufficient amount of sample.
3. Place the test pen vertically into the extraction buffer container, gently press to break the foil. The edge of the extraction buffer container should align with the top edge of the marking ring on the test device.
4. Read the result after 15 minutes.
IMPORTANTLY:
a) It is necessary to obtain as much sample as possible
b) This procedure may cause discomfort. Do not advance the pad further if there is strong resistance. Testing of children aged 2 to 15 years should be performed by an adult.
Attention! The nasal swab pen contains two test strips:
1 - СOV - for diagnosing COVID-19 virus antigen (blue)
2 - FLU - for the diagnosis of influenza A/B virus antigen (green)
IMPORTANTLY
Care must be taken to ensure that the marking ring on the test device aligns with the edge of the extraction buffer container.
Otherwise, it will lead to lateral flow disruption and invalid test result.
INTERPRETATION OF TEST RESULTS
POSITIVE
For COVID-19 test. COV test strip
Two colored lines appear on the membrane. One line appears in the control region (C) and the other line appears in the test region (T).
For influenza A test. FLU test strip
One colored line appears in the control region (C). A second line appears in the test region A (at the lower edge).
For influenza B test. FLU test strip
One colored line appears in the control region (C). A second line appears in the test region B (inside the membrane).
For testing for influenza A and B. FLU test strip
One colored line appears in the control region (C). Two lines appear in the test region.
Caution! Co-infection with influenza A and B is rare. If the test shows a positive result for influenza A and B, it must be repeated. If the test shows such a result again, it is necessary to test with a different method.
NEGATIVE
For COVID-19 test. COV test strip
Only one colored line appears in the control region (C).
No visible colored lines appear in the test region (T).
For testing for influenza A and B. FLU test strip
Only one colored line appears in the control region (C).
No visible colored bands appear in the test region (B) and (A).
INVALID
Any test result that does not produce a line in the control region after the specified reading time is invalid. Please review the test procedure and repeat it using a new test. If the problem persists, discontinue use of the kit immediately and contact your local distributor.
NOTE:
1. The intensity of the color of the strip or strips in the test region (T), (A), (B) may vary depending on the concentration of the analytes present in the sample. Therefore, the appearance of any shade of color in the test region should be considered a positive result. Please note that this is only a qualitative test, so it cannot determine the concentration of the analytes in the sample.
2. Insufficient sample volume, incorrect test procedure, or using the test after the expiration date are the most likely causes of an incorrect test result.
Application features
Internal procedural control:
MILISTAN COMBI nasal test Express test for detection of COVID-19 virus antigen and influenza A/B has a built-in (procedural) control. Each strip on the test device has an internal standard zone to ensure proper movement of samples through the membrane.
Before reading the result, the user must confirm the presence of a colored band located in the control zone “C”.
External positive and negative controls:
According to standards of laboratory practice, external positive and negative controls can be used to verify correct test performance.
LIMITATIONS OF THE TEST METHOD
1. MILISTAN COMBI nasal test The rapid test for the detection of COVID-19 virus antigen and influenza A/B is intended for in vitro diagnostics and should be used only for the qualitative detection of SARS-CoV-2, influenza A and influenza B virus antigens. The color intensity of the positive result strip should not be evaluated as “quantitative or semi-quantitative”.
2. The rapid test detects both viable and non-viable viruses.
3. As with all diagnostic tests, the final clinical diagnosis should not be based on the results of a single test, but should be made by a physician only after evaluating all clinical and laboratory studies.
4. Failure to follow the TEST PROCEDURE and INTERPRETATION OF RESULTS may adversely affect the effectiveness of the test and/or lead to an erroneous test result.
5. The results obtained with this assay, especially in the case of test strips with weak coloration that are difficult to interpret, should be used in conjunction with other available clinical information. In any case, a physician should be consulted.
6. Negative test results do not exclude viral infection and must be confirmed by molecular analysis.
PERFORMANCE INDICATORS
The limit of detection (LOD) of the MILISTAN COMBI nasal test Rapid test for the detection of COVID-19 and influenza A/B virus antigen, defined as the concentration of influenza virus and SARS-CoV-2 virus that gives a positive test result, is approximately 95%, as determined by evaluating different concentrations of inactivated influenza A virus (H3N2, H1N1), inactivated influenza B virus (Victoria, Yamagata) and inactivated SARS-CoV-2 virus with the MILISTAN COMFORT nasal test Rapid test for the detection of COVID-19 and influenza A/B virus antigen. 20 tests were performed for each concentration. The test results determined a concentration of 1.0 × 104 TCID50/mL as the LOD for influenza A (H3N2), 4.3 × 104 TCID50 as the LOD for influenza A (H1N1), 2.2 × 105 TCID50 for influenza B (Victoria), 2.5 × 105 TCID50 for influenza B (Yamagata), and 1 × 102.4 TCID50/mL as the LOD for SARS-CoV-2.
Clinical assessment:
To detect COVID-19 virus antigen:
A total of 197 clinical samples were selected to test the effectiveness of the MILISTAN COMBI nasal test Express test for the detection of COVID-19 virus antigen and influenza A/B in comparison with RT-PCR testing. No differences in test results were found depending on the age and gender of the patient. 41 samples were detected positive by RT-PCR, and 156 samples were negative when tested by RT-PCR. These samples were tested using the MILISTAN COMBI nasal test Express test for the detection of COVID-19 virus antigen and influenza A/B. The results are shown in Table 1.
Table 1: Results of testing with the Rapid Test for the Detection of COVID-19 Virus Antigen in Comparison with RT-PCR Testing
| 3T-PCR |
| Positive | Negative | Total |
| Rapid test for detection of COVID-19 virus antigen | Positive | 39 | 1 | 40 |
| Negative | 2 | 155 | 157 |
| Total | 41 | 156 | 197 |
Relative sensitivity: 95.1% (83.9%99.3%) *
Relative specificity: 99.4 % (96.5%99.9%)*
Overall agreement of results: 98.5% (95.6%99.5%)*
*95% confidence interval
For the detection of A/B virus antigens:
A total of 197 collected samples were tested with the MILISTAN COMBI nasal test Rapid test for the detection of COVID-19 virus antigen and influenza A/B and these results were compared with the RT-PCR method.
For all these samples, the overall sensitivity of the Rapid Test for the detection of influenza A/B virus antigen compared to RT-PCR was 95.2% (60/63) for influenza A and 93.1% (54/58) for influenza B. The overall specificity was 100.0% (134/134) for influenza A and 100.0% (139/139) for influenza B. The results are presented in Tables 2 and 3.
Table 2: Results of testing with the Rapid Influenza A Virus Antigen Detection Test compared to RT-PCR testing
| 3T-PCR |
| Positive | Negative | Total |
| Rapid test for detection of influenza A virus antigen | Positive | 60 | 0 | 60 |
| Negative | 3 | 134 | 137 |
| Total | 63 | 134 | 197 |
Relative sensitivity: 95.2 %( 86.9%98.4%) *
Relative specificity: 100.0 %(97.2%100.0%) *
Overall agreement of results: 98.5%( 95.6%99.5%) *
*95% confidence interval
Table 3: Results of testing with the Rapid Influenza B Antigen Test compared to RT-PCR testing
| 3T-PCR |
| Positive | Negative | Total |
Rapid test for detection of influenza B virus antigen | Positive | 54 | 0 | 54 |
| Negative | 4 | 139 | 143 |
| Total | 58 | 139 | 197 |
Relative sensitivity: 93.1 %( 83.6%97.3%) *
Relative specificity: 100.0 %( 97.3%100.0%) *
Overall agreement of results: 98.0 %( 94.9%99.2%) *
*95% confidence interval
Cross-reactivity:
Cross-reactivity with the following organisms was investigated. Samples positive for the following organisms were considered negative when tested with the MILISTAN COMBI Nasal Rapid Test for COVID-19 and Influenza A/B Antigen.
| HCoV-229E | Bordetella para pertussis | Adenovirus 1 |
| HCoV-OC43 | Bordetella pertussis | Adenovirus 2 |
| HCoV-NL63 | Candida albicans | Adenovirus 3 |
| MERS-coronavirus | Chlamydia pneumoniae | Adenovirus 4 |
| SARS-coronavirus | Group C streptococcus | Adenovirus 5 |
| Human metapneumovirus | Haemophilus influenzae | Adenovirus 7 |
| Norovirus | Legionella pneumophila | Adenovirus 55 |
| Parainfluenza virus 1 | Mycoplasma pneumoniae | Epstein-Barr virus |
| Parainfluenza virus 2 | Mycobacterium tuberculosis | Enterovirus EV70 |
| Parainfluenza virus 3 | Staphylococcus aureus | Enterovirus EV71 |
| Parainfluenza virus 4 | Staphylococcus epidermidis | Enterovirus A16 |
| Respiratory syncytial virus | Stregacoccus agalactiae | Enterovirus A24 |
| Rhinovirus A30 | Streptococcus pneumoniae | Enterovirus B1 |
| Rhinovirus B52 | Streptococcus pyogenes | Echovirus 6 |
The following substances, naturally present in respiratory specimens or which may be introduced into the respiratory tract, were evaluated at the following concentrations as listed below. None of them had any effect on the test results using the MILISTAN COMBI Nasal Test Rapid Test for the Detection of COVID-19 and Influenza A/B Virus Antigen.
| Substance | Concentration | Substance | Concentration |
| Nasal sprays | 10% | Guaiacol glycerol ester | 20 mg/mL (mg/ml) |
| Mouthwashes | 10% | Mucin | 1% |
| Throat sprays | 10% | Whole blood | 4% |
| 4-acetamidophenol | 10 mg/mL (mg/ml) | Mupirocin | 250 µg/mL (μg/mL) |
| Acetylsalicylic acid | 10 mg/mL (mg/ml) | Oxymetazoline | 25 µg/mL (μg/mL) |
| Albuterol | 10 mg/mL (mg/ml) | Phenylephrine | 10 mg/mL (mg/ml) |
| Chlorpheniramine | 5 mg/mL (mg/ml) | Phenylpropanolamine | 1 mg/mL (mg/ml) |
| Dexamethasone | 50 µg/mL (μg/mL) | Zanamivir | 10 mg/mL (mg/ml) |
| Dextromethorphan | 10 µg/mL (μg/mL) | Adamantanamine | 500 ng/mL (ng/ml) |
| Diphenhydramine | 5 mg/mL (mg/ml) | Oseltamivir phosphate | 10 mg/mL (mg/ml) |
| Doxylamine succinate | 1 mg/mL (mg/ml) | Tobramycin | 10 mg/mL (mg/ml) |
| Flunisolide | 25 µg/mL (μg/mL) | Triamcinolone | 14 mg/mL (mg/ml) |
Producer
Ashur Tek. (Hangzhou) Co., Ltd. / Assure Tech. (Hangzhou) Co., Ltd.
Building 4, No.1418-50, Moganshan Road, Gongshu District, Hangzhou, 310011 Zhejiang, PRChina / Building 4, No.1418-50, Moganshan Road, Gongshu District, Hangzhou, 310011 Zhejiang, PRChina
Authorized representative:
Mili Healthcare Limited 01032, Ukraine, Kyiv, Shevchenkivskyi district, T. Shevchenko boulevard, Building 33B, BS5
