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Kit for detecting novel coronavirus and amplification primer composition thereof

A coronavirus and amplification primer technology, which is applied in the field of inspection and quarantine, can solve the problems of poor accuracy, sensitivity, and repeatability of the kit, and insufficient performance and verification of the reagent, so as to reduce false negatives and missed detection, solve Effects of insufficient coverage and high reliability of results

Active Publication Date: 2021-01-08
海关总署(北京)国际旅行卫生保健中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The new type of coronavirus pneumonia is a public health emergency. All reagent manufacturers have completed the development and approval of nucleic acid detection kits in the shortest possible time. Most of the currently used nucleic acid detection kits have not yet undergone sufficient clinical evaluation, and the approved reagents have not been adequately tested. The performance verification of diagnostic kits from different manufacturers is inconsistent, the detection capabilities are uneven, and the accuracy, sensitivity, and repeatability of some kits are not good.
[0009] Therefore, the existing novel coronavirus detection methods still need to be improved

Method used

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  • Kit for detecting novel coronavirus and amplification primer composition thereof
  • Kit for detecting novel coronavirus and amplification primer composition thereof
  • Kit for detecting novel coronavirus and amplification primer composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Using the kits and testing procedures described in the general method, clinical samples were verified and tested in the Beijing port laboratory, Customs A, Customs B, Customs C, and disease control confirmation experiments.

[0070] sample source

[0071] The test samples came from the throat swab samples of inbound personnel at Beijing Port, A Customs, B Customs, and C Customs laboratories. There were 23 samples from Beijing Port, of which 16 were positive and 7 were negative; A Customs had 26 samples, of which 10 were positive. 16 were negative; 36 were from customs B, of which 19 were positive and 17 were negative; 21 were from customs C, of ​​which 4 were positive and 17 were negative; Pharyngeal swab samples of personnel, of which 16 were positive and 16 were negative.

[0072] Materials and Methods

[0073] The following kits and equipment were used for detection:

[0074] New Coronary Pneumonia Virus (SARS-CoV-2) nucleic acid multiplex real-time fluorescent RT...

Embodiment 2

[0081] According to the embodiment of the present invention, the inventor also verified the quantitative ability of the kit of the present invention, wherein, Figure 2 shows that according to an embodiment of the present invention, the plasmid standard (concentration is 3.1 × 10 10 copies / μl) for a 10-fold serial dilution amplification curve, figure 1 shows a standard curve according to one embodiment of the present invention, the concentration of the plasmid standard used is 3.1 × 10 10 copy / microliter, carry out 10-fold serial dilution on the plasmid standard, and the copy number of the plasmid standard is 3.1×10 7 ~3.1×10 3 For copies / microliter, the Ct values ​​are: 10.91, 14.34, 17.92, 21.32 and 24.62, respectively. Therefore, there is a good logarithmic linear relationship between the logarithmic value of the copy number of the plasmid standard and the Ct value. Specifically, the slope of the standard curve of the real-time fluorescent quantitative PCR reaction system ...

Embodiment 3

[0083] According to the embodiments of the present invention, the inventor has further verified the specificity of the kit of the present invention, image 3 Shows the amplification curve according to one embodiment of the present invention, it can be seen from the fluorescence amplification curve that influenza A virus, influenza B virus, Norovirus, HIV virus nucleic acid, Zika virus, dengue virus, measles The amplification curves of virus samples such as virus, yellow fever virus, and hepatitis C virus and the negative control did not show typical amplification curves, but only the new coronavirus showed typical amplification curves.

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Abstract

The present invention proposes a set of amplification primer composition for novel coronavirus and a kit using the amplification primer composition, the amplification primer composition includes: a first primer set, the first primer set is suitable for specifically amplifying the 3'-UTR target region; a second primer set adapted to specifically amplify the E gene target region; and a third primer set adapted to specifically amplify Increase N gene target region. The amplification primer composition can efficiently and simultaneously detect novel coronaviruses.

Description

technical field [0001] The invention relates to the field of inspection and quarantine, in particular, the invention relates to an amplification primer composition for novel coronavirus and a fluorescent multiplex RT-PCR kit for detecting novel coronavirus. Background technique [0002] New coronary pneumonia is mainly transmitted through respiratory droplets and close contacts, and there is a possibility of aerosol transmission when exposed to high-concentration aerosols for a long time in a relatively closed environment. Most patients will have clinical manifestations such as fever, dry cough, and fatigue. Severe patients often develop dyspnea and / or hypoxemia one week after the onset, and severe cases can rapidly progress to acute respiratory distress syndrome. The multiple transmission routes of COVID-19 lead to rapid transmission, and patients are prone to "cytokine storm", which is ultimately life-threatening, especially for the elderly and those with chronic underlyin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2563/107C12Q2537/143C12Q2521/107C12Q2561/113Y02A50/30
Inventor 刘翌肖利力
Owner 海关总署(北京)国际旅行卫生保健中心