inquiry@rdetect.com 【SUMMARY】
The novel coronaviruse belongs to the β genus. COVID-19 is an acute respiratory infectious disease which people are generally susceptible to. Currently, patients infected by the novel coronavirus are the main source of infection; the asymptomatic infected can also be a source of infection. Based on the current epidemiological investigation, the incubation period of COVID-19 is between 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 also found in cases.
Influenza is a highly contagious, acute, viral infection of the respiratory tract. It is an infectious disease easily spreading through coughing and sneezing droplets containing live viruses. Type A viruses are typically more prevalent than type B viruses and are associated with most serious influenza epidemics, while type B infections are usually milder. Rapid diagnosis of influenza A/B has become more important due to the availability of effective antiviral therapy. Rapid diagnosis of influenza can reduce hospital stays, antimicrobial use and cost of hospital care.
【PRINCIPLES OF TEST】
Anti-SARS-CoV-2 antibody is coated in the test region of SARS-CoV-2. Anti-Influenza A antibody and anti-Influenza B antibody are coated in the test A region (A) and test B region (B) respectively. During testing, antigen in the specimen reacts with anti-SARS- CoV-2 antibody-coated particles and with anti-Influenza A antibody-coated particles as well as with anti-Influenza B antibody-coated particles in the reaction pad to produce the immune complex. The complex migrates along the membrane by capillary action to the test region. The complex then respectively reacts with the anti-SARS-CoV-2 antibody in the test region of SARS-CoV-2 and with anti- Influenza A antibody in the A region as well as with anti-Influenza B antibody in the B region. If the specimen contains SARS-CoV-2 antigen, a colored line will appear in the test region of SARS-CoV-2. If the specimen contains Influenza A and/or Influenza B viruses, a colored line will appear in the test A and/or B region. If the specimen does not contain antigen of SARS-CoV-2, Influenza A and/or Influenza B viruses, no colored line will appear in the test region, indicating a negative result. To serve as a control, a colored line will always appear in the control region (C), indicating that the result is effective.
【SPECIMEN COLLECTION AND HANDLING】
- Nasopharyngealswabprocedure: As shown in the figure below, carefully and slowly insert the sterile swab into the nostril carefully. Gently push the swab until meeting resistance at the wall of the turbinate. Rotate the swab gently against the nose wall for a few times. If coughing, please wait for a while and try again. Don’t push/pull the swab hard to avoid injuring the tester.
- Oropharyngealswabprocedure: As shown in the figure below, after rinsing with saline, insert the sterile swab into the oral pharynx, and gently rub the back wall of the pharynx and tonsils for several times. Don’t push/pull the swab hard to avoid injuring the tester.
【OPERATING STEPS】Before testing, read the operating instruction carefully, and restore the testing kit and samples to room temperature (20-25℃).- Nasopharyngeal/oropharyngealspecimen
STEP 1: Tear off the sealing film of the extraction tube.
STEP 2: Dip the swab into the extraction tube (the solution should fully permeate the swab). Rotate and squeeze the swab at least 10 times.
STEP 3: Remove the swab and cover the tube with the fitting.
STEP 4: Open the foil bag, take out the test card, place on a horizontal table and use it as soon as possible within 15 minutes. Vertically drop 3 drops (about 100μL) of the treated sample solution into the sampling hole of the test card.
STEP 5: The test card is kept at room temperature (20-25℃) for 15 minutes to read the results, but the results over 30 minutes were invalid.
【INTERPRETATION OF RESULT】
【LIMITATION】- Blood,serum,plasma, urine, stool and other samples may cause abnormal results.
- Positiveresultdoes not rule out infections with other pathogens.
- Negative resultmaybe caused by improper specimen collection and transfer, or by low concentration of antigen from the specimen. The specimen should be further analyzed to confirm the cause.
- Aswithall diagnostic tests, all results must be interpreted together with other clinical information available to the physician.
- Inconsistentandincorrect result may be caused by improper technical and procedural operation. Specimen contaminated and hemolyzed and other drugs may interfere with the testing resul.
【PERFORMANCE】
- Positive reference coincidence rate: the positive reference coincidence rate of the enterprise should be 100%.
- Negative reference product conformity rate: the negative reference product conformity rate of the enterprise should be 100%.
- Limit of detection (LoD)：
① The detection limit of SARS-CoV-2 is 40 TCID50/mL.
② The detection limit of Influenza A
Type Estimated LOD Flu A No. Positive/Total % Positive Flu A/California/7/2009(H1N1) 7.8×103 TCID50/L 20/20 100% FluA/New Caledomia/20/99(H1N1) 9.0×103 TCID50/L 19/20 95% Flu A/Brisbane/59/2007(H1N1) 2.8×104TCID50/L 20/20 100% FluA/GuangdongMaonan/SWL1536/2019(H1N1) 2.2×104 TCID50/L 20/20 100% Flu A//Uruguay/716/2007(H3N2) 1.1×103TCID50/L 20/20 100% FluA/New York/55/2004 (H3N2) 9.0×102 TCID50/L 20/20 100% Flu A/Texas/50/2012(H3N2) 1.1×103 TCID50/L 19/20 95% Flu A/pigeon/Zhejiang/112089/2014 (H5N1) 2.5×104 EID50/ml 20/20 100% Flu A/chicken/Zhejiang/131146/2016 (H7N9) 1.2×104 EID50/ml 20/20 100% Flu A/chicken/Zhejiang/329/2011 (H9N2) 1.6×104 EID50/ml 20/20 100% ③ The detection limit of Influenza B
Type Estimated LOD Flu B No. Positive/Total % Positive FluB/Washington/02/2019(Victoria) 1.4×105TCID50/L 20/20 100% FluB/Malaysia/2506/04(Victoria) 1.1×105TCID50/L 19/20 95% FluB/Brisbane/60/2008(Victoria) 3.5×105TCID50/L 20/20 100% FluB/Phuket/3073/2013 (Yamagata) 7.0×103TCID50/L 20/20 100% - Cross-reactivity
① Cross reactivity of SARS-CoV-2
When the concentration of novel coronavirus is between 1.0 × 102 and 3.0 × 105 TCID50/mL, there is no cross-reactivity with influenza A virus and influenza B virus antigen tests.
② Cross reactivity of influenza A virus and influenza B virus
The following influenza A virus strains, within the concentration range of 1.0 × 103-1.0 × 106 TCID50/L, will not cross-react with antigen tests for COVID-19 and influenza B virus: Flu A/California/7/2009(H1N1), Flu A/New Caledomia/20/99(H1N1), Flu A/Brisbane/59/2007(H1N1), Flu A/GuangdongMaonan/SWL1536/2019(H1N1), Flu A//Uruguay/716/2007(H3N2), Flu A/New York/55/2004 (H3N2), and Flu A/Texas/50/2012(H3N2).The following influenza A virus strains, within the concentration range of 1.0×105-1.0×106 EID50/ml, will not cross-react with antigen tests for COVID-19 and influenza B virus: Flu A/pigeon/Zhejiang/112089/2014 (H5N1), Flu A/chicken/Zhejiang/131146/2016 (H7N9), Flu A/chicken/Zhejiang/329/2011 (H9N2).
The following influenza B virus strains, within the concentration range of 1.0 × 104-5.0 × 106 TCID50/L, will not cross-react with antigen tests for COVID-19 and influenza A virus: Flu B/Washington/02/2019(Victoria), Flu B/Malaysia/2506/04(Victoria), Flu B/Brisbane/60/2008(Victoria), and Flu B/Phuket/3073/2013 (Yamagata).
③ Cross reactivity of other pathogens
12 types of viruses (1.0×105TCID50/mL), 8 types of bacteria (1.0×106Pfu/mL), Mycoplasma pneumoniae (1.0×106Pfu/mL) and Chlamydia pneumoniae (1.0×106Pfu/mL) listed below are confirmed to have no cross-reactivity with SARS-CoV-2 & Flu A/B Antigen Rapid Test Kit.
Human coronavirus 229E Human coronavirus OC43 Human coronavirus NL63 Human Metapneumovirus (HMPV) Adenovirus Respiratory syncytial virus Rhinovirus Enterovirus Parainfluenza virus l-4 Haemophilus influenzae Chlamydia pneumoniae Mycoplasma pneumoniae Legionella pneumoniae Bordetella pertussis Staphylococcus aureus Streptococcus pneumoniae Staphylococcus epidermidis Streptococcus pyogenes Candida albicans - Interference
No interfering is observed with interference substances listed below at the indicated concentration.
Substance Concentration Benzocaine 150mg/dL Blood 5% Mucin 5 mg/mL Naso GEL(NeilMed) 5% Phenylephrine 15% Afrin Oxymetazoline 15% CVS Nasal Spray (Cromolyn) 15% Alkalol 10% Sore Throat Phenol Spray 15% Tobramycin 3.3mg/dL Mupirocin 0.15mg/dL Fluticasone 0.000126mg/dL Tamiflu (Oseltamivir phosphate) 500mg/dL Budenoside 0.00063 mg/dL Biotin 0.35mg/dL Methanol 150mg/dL Acetylsalicylic Acid 3mg/dL Diphenhydramine 0.074mg/dL Dextromethorphan 0.00156mg/dL Dexamethasone 1.2mg/dL Mucinex 5% - Clinicalperformance
① Results and Analysis of SARS-CoV-2
Novel Coronavirus (2019-nCoV) Nucleic Acid Test kit Total Positive Negative Positive 99 0 99 Negative 4 300 304 Total 103 300 403 Statistic Value 95% Cl Sensitivity 96.12% 90.35% to 98.93% Specificity 100% 98.78% to 100.00% Total coincidence rate 99.01% 97.48% to 99.73% The 95% confidence intervals of sensitivity, specificity, the total coincidence rate is calculated based on the binomial distribution. Analysis results by Kappa consistency test shows that the Kappa=0.97. (As Kappa>0.75, it suggests good consistency between the candidate kit and the reference kit.
The results correlated with the Ct value of the positive samples from 103 Confirmed SARS-CoV-2 Cases were as follows:
Ct Value Number of cases SARS-CoV-2 Ag Test positive SARS-CoV-2 Ag Test negative Sensitivity PCR positive 103 99 4 96.12% Ct≤30 68 67 1 98.52% Ct≤33 98 95 3 96.93% Ct≤35 103 99 4 96.12% ② Results and Analysis of Influenza A
Influenza A and B Virus Nucleic Acid Test kit Total Positive Negative Positive 49 0 49 Negative 4 300 304 Total 53 300 353 Statistic Value 95% Cl Sensitivity 92.45% 81.79% to 97.91% Specificity 100% 98.78% to 100.00% Total coincidence rate 98.87% 97.12% to 99.69% The 95% confidence intervals of sensitivity, specificity, the total coincidence rate is calculated based on the binomial distribution. Analysis results by Kappa consistency test shows that the Kappa=0.95. (As Kappa>0.75, it suggests good consistency between the candidate kit and the reference kit.
The results correlated with the Ct value of the positive samples from 53 Confirmed FluA Cases were as follows:
Ct Value Number of cases Flu A AgTest positive Flu A Ag Test negative Sensitivity PCR positive 53 49 4 92.45% Ct≤30 20 20 0 100% Ct≤33 50 47 3 94% Ct≤35 53 49 4 92.45% ③ Results and Analysis of Influenza B
Influenza A and B Virus Nucleic Acid Test kit Total Positive Negative Positive 56 0 56 Negative 2 300 302 Total 58 300 358 Statistic Value 95% Cl Sensitivity 96.55% 86.09% to 99.58% Specificity 100% 98.78% to 100.0% Total coincidence rate 99.44% 98.00% to 99.93% The 95% confidence intervals of sensitivity, specificity, the total coincidence rate is calculated based on the binomial distribution. Analysis results by Kappa consistency test shows that the Kappa=0.98. (As Kappa>0.75, it suggests good consistency between the candidate kit and the reference kit.
The results correlated with the Ct value of the positive samples from 58 Confirmed FluB Cases were as follows:
Ct Value Number of cases Flu B AgTest positive Flu B Ag Test negative Sensitivity PCR positive 58 56 2 96.55% Ct≤30 12 12 0 100% Ct≤33 49 48 1 94% Ct≤35 58 56 2 96.55% - Highdosehook effect
No high dose hook effect was observed up to SARS-CoV-2 at a concentration of 3.0 x 105 TCID50/mL, Flu A and Flu B at concentrations of 1.0 × 106 TCID50/L and 5.0 × 106 TCID50/L, respectively.
- Repeatability
The repeatability reference products of the enterprise were tested, repeated for 10 times, and the positive coincidence rate is 100%.
