SARS-CoV-2 Nucleic Acid Detection Fluorecent Kit rtPCR CE

Intended use
For in vitro qualitative detection of SARS-CoV-2 (also known as 2019-nCoV) ORF1ab, E, and N gene in oropharyngeal swabs and sputum specimens of suspected SARS-CoV-2-infected pneumonia cases, suspected clustered infection cases, and others who need SARS-CoV-2 infection diagnosis or differential diagnosis.

Test principle
This product is designed as a multiplex real-time reverse-transcription PCR system, containing specific primers and fluorescent probes targeting ORF1ab, E and N gene of SARS-CoV-2. Virus nucleic acid is detected by monitoring fluorescence intensity in real-time. Internal control is added to monitor the presence of PCR inhibitor within specimen, thus effectively prevent false negative results.
Reagents components
Component Ingredient 96 Tests
qRT-PCRReaction Mix Primer, probe, dNTP, Mg2+, buffer 816 μL×2 μL×1
qRT-PCR Enzyme Mix Taq polymerase, Uracil-DNA glycosylase 288 μL×1
Negative Control DEPC-treated water 1350μL×1
Positive Control Armored RNA containing target gene fragment 1350 μL×1
Internal Control Armored RNA containing Internal Control gene fragment 192μL×1

Please read this manual carefully before use. Please follow the procedures below.

Performance characteristics

1. Coincidence rate of negative reference materials
Detection of national negative reference materials, the results are all negative; or test the enterprise negative reference materials, and the coincidence rate of negative reference materials (-/-) is 10/10.

2. Coincidence rate of positive reference materials
Detection of national positive reference materials, the results meet the requirements of the national reference materials; or test the enterprise positive reference materials, and the coincidence rate of positive reference materials (+/+) is 5/5.

3. Repeatability
Detection of national repeatability reference materials, the repeatability conforms to the coefficient of variation (CV) ≤5%; or test the enterprise repeatability reference materials, the repeatability within the batch conforms to CV≤5%

4. Limit of detection (LOD)
Detection of national sensitivity reference materials, S1-S3 are positive; or test three enterprise sensitivity reference materials, S1 and S2 are detected at least, that is, the LOD is not higher than 1×103 copies/mL.

5. Cross-reactivity
The product shows no cross-reactivity with positive samples of human coronaviruses (HKU1，OC43，NL63 and 229E), HCoV-SARS, HCoV-MERS;H1N1, seasonal H1N1, seasonal H3N2, seasonal H5N1, seasonal H7N9, B/Yamagata, B/Victoria, Respiratory syncytial virus types A and B, Parainfluenza virus types 1, 2, and 3, Rhinovirus types A, B, C, Adenovirus types 1, 2, 3, 4, 5, 6, 7, 55, Enterovirus types A, B, C, D, Human lung virus (HPVV), EB virus, Measles virus, Human cytomegalovirus, Rotavirus, Norwalk viruses, Mumps virus, Chickenpox-zoster virus; Mycoplasma pneumoniae, Chlamydia pneumoniae; Legionella, Bacillus pertussis, Haemophilus influenza, Staphylococcus aureus, Pyogenic streptococcus, Streptococcus pneumoniae, Klebsiella pneumonia, Mycobacterium tuberculosis; Aspergillus fumigatus, Candida albicans, Canadida glabrata, cryptococcus neoformans, Pseudomonas aeruginosa, and human genomic DNA.

6. Interfering substances
0.9 g/mL sodium chloride (including 1% PC-300), 100μg/mL phenylephrine, 100μg/mL oxymetazoline, 100μg/mL beclomethasone, 100μg/mL flunisolide, 100μg/mL triamcinolone acetonide, 200μg/mL budesonide, 200μg/mL mometasone, 200μg/mL fluticasone propionate, 200μg/mL histamine hydrochloride, 10 mg/mL ribavirin, 400 U/μL α-interferon, 0.1mg/mL dexamethasone, 0.568μg/mL zanamivir, 100μg/mL oseltamivir, 100μg/mL peramivir, 100μg/mL lopinavir, 100μg/m ritonavir, 100μg/mL arbidol, 100μg/mL levofloxacin, 200μg/mL azithromycin, 100μg/mL ceftriaxone, 100μg/mL meropenem, 100μg/mL tobramycin, 0.25 g/L mucin and 5% (V/V) whole blood have no significant effect on the test results of the kit.

7. Clinical research

This product was compared with similar products approved by the NPMA for clinical comparison studies on 537 samples in 4 clinical institutions. The sample types verified in the clinical trials were oropharyngeal swabs and sputum, and some samples were verified using the stored nucleic acid extract. The total coincidence rate was 99.44%, the negative coincidence rate was 99.35%, and the positive coincidence rate was 99.56%. Further clinical data will be collected to validate the clinical performance of the product after the launch.

Storage and stability
The reagents are stable until the expiration dates labelled on the package, when stored upright at -10°C ~-30°C and sealed. Protect from direct sunlight. Freeze-thaw cycle should not exceed 6 times.