2019-nCoV Molecular Diagnostic Kit (Fluorescent PCR Test)

【Test Principle】

In this kit, primers and probes of real-time fluorescent PCR technology are designed to the conserved and specific regions of the ORF1ab and N genes of 2019-nCoV, respectively. During PCR amplification, the probe binds to the template, and the 5'-end reporter group of the probe is cleaved by the Taq enzyme (5'→3' exonuclease activity), thereby moving away from the quenching group to generate a fluorescent signal. The real-time amplification curve is automatically plotted based on the detected fluorescence signal, and the sample Ct value is calculated. FAM and HEX fluorophores are labeled to ORF1ab gene and N gene probes in 2019-nCoV-PCR reaction solution. By using one test, qualitative detection of the above two genes of 2019-nCoV can be performed simultaneously.The kit is provided with an internal control (IC), which is a human housekeeping gene fragment, and is labeled with a ROX fluorescent group.

Note: The different lots of components cannot be exchanged .

【Storage conditions and expiration date】

The kit is transported at room temperature.

For long-term preservation, the kit should be stored in the dark at

-25℃～-15℃ and is valid for 12 months.

Date of manufacture and expiry date (see label).

【Sample requirements】

1. Applicable samples Nasopharyngeal swab
2. Samples collection Nasopharyngeal swab: the nasopharyngeal secretions were swabbed with sterile swab and placed in the tube containing the preservation solution.
3. Sample handling and preservation:The samples used for nucleic acid detection shall be tested as soon as possible. If the samples can be tested within 24 hours, they could bestored at 2～8℃; the samples should be stored at -70℃ or below (if there is no -70℃ storage condition, they shall be temporarily stored in -20℃). Repeated freezing and thawing shall be avoided during sample transportation.

   【Test method】

     Nucleic Acid Extraction
     1.1 Manual nucleic acid extraction Negative Control, add same volume of sample nucleic acid extraction and purification reagent to the tube (sample and nucleic acid extraction and purification reagen 1:1 for extraction).PCR reactions for nucleic acid amplification.
   

    1.2 Machine Extraction of Nucleic Acid:

   This kit is compatible with any other commercial kits. For other commercial nucleic acid extraction kits, please follow the instructions. 
   
   System preparation and Loading
   

   When start using 2019-nCoV-PCR Reaction Solution and RT-PCR Enzyme Mix, add Rehydration (sterilized purified water)1250 μL/tube and 250 μL/tube to dissolve respectively. This is done by vortex for 20 seconds followed by a brief spin.Preparation of 2019-nCoV-RT-PCR Mixture: take n×25 μL 2019-nCoV-PCR reaction solution and n×5 μL Enzyme Mix (n as the number of reaction tubes), mixture and centrifuge. And each tube was dispensed in an amount of 30μL in a PCR tube suitable for a fluorescence PCR apparatus. 20μL of Negative Control, the extracted sample nucleic acid and 2019-nCoV-PCR Positive Control are added to reaction systems, and the total reaction volume is 50μL. Fasten the tube cover and move it to the amplification test area after a few seconds of vortex mix and centrifugation.
   

NOTE:The remaining reagent should be stored below -15℃～-25℃ ,frozen and thawed repeatedly for no more than 5 times.Usen immediately after mixing 2019-nCoV-PCR Reaction solution and RT-PCR Enzyme Mix. Discard the extra.

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FAQ

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