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Reaction system for detecting low-abundance novel coronavirus and method and application thereof

A reaction system, coronavirus technology, applied in biochemical equipment and methods, microbial determination/inspection, resistance to vector-borne diseases, etc., can solve problems affecting prevention and control, and achieve stable detection rate and high sensitivity , the effect of reducing cross-contamination

Active Publication Date: 2020-08-21
北京元码医学检验实验室有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although various methods for nucleic acid detection have been developed, various false negatives are still unavoidable. These false negative cases have brought a very adverse impact on the prevention and control of this highly contagious disease, seriously affecting the prevention and control , prevention

Method used

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  • Reaction system for detecting low-abundance novel coronavirus and method and application thereof
  • Reaction system for detecting low-abundance novel coronavirus and method and application thereof
  • Reaction system for detecting low-abundance novel coronavirus and method and application thereof

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Embodiment

[0055] 1. According to the genome sequence information of the new coronavirus (download the complete reference genome sequence of the virus (GWHABKF00000000, GWHABKG00000000, GWHABKH00000000, GWHABKI00000000, GWHABKJ00000000, etc.) N gene consensus sequence segment) to design primers for the first amplification reaction and primers / probes for the second amplification reaction (qPCR). The primers for the first amplification reaction include two outer primers (F3, B3), two inner primers (FIP, BIP) and two loop primers (LF, LB). The specific sequences of the primers are shown in the table below, and the positions of the primers see relation Figure 4 .

[0056] Primer / Probe Name sequence SEQ ID No: qPCR-F GGGGAACTTCTCCTGCTAG 7 qPCR-R CAGACATTTTGCTCTCAAG 8 qPCR-P TTGCTGCTGCTTGACAG 9 F3 primer AGCCTCTTCTCGTTCCTC 3 B3 primer AGTGACAGTTTGGCCTTG 4 FIP primer GCCAGCCATTCTAGCAGGAGAACACGTAGTCGCAACAGT 1 BIP primer GCGGT...

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Abstract

The invention discloses a reaction system, a method and an application for detecting low-abundance novel coronavirus. An isothermal amplification reaction and a PCR amplification reaction are carriedout in the same reaction system, mutual influence between the isothermal amplification reaction and the PCR amplification reaction is avoided, and the purpose of detecting the low-abundance nucleic acid is achieved. The lower limit of the method is lower than that of a single PCR detection method by one order of magnitude or above, the detection sensitivity and specificity are further improved, the detection rate is stabilized, and meanwhile quantitative analysis of the virus load is conducted through a qPCR amplification curve CT value. Through the one-tube method design of integrating specific signal amplification of two groups of reactions and high-sensitivity fluorescence detection, the detection time is shortened to be within half an hour and is shortened by more than three times compared with conventional detection, so that the purpose of quickly and stably detecting extremely low abundance pathogens is achieved, and the problem of missing detection in trace pathogen sample detection is prevented.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a reaction system, method and application for detecting low-abundance novel coronaviruses. Background technique [0002] At present, nucleic acid detection is the most accurate and convenient way to diagnose new coronavirus infection. Although various methods for nucleic acid detection have been developed, various false negatives are still unavoidable. These false negative cases have brought a very adverse impact on the prevention and control of this highly contagious disease, seriously affecting the prevention and control , Prevention and treatment. Contents of the invention [0003] To solve at least some of the technical problems in the prior art, the present invention provides a reaction system for detecting low-abundance novel coronaviruses in one tube, and based on this, the present invention is completed. Specifically, the present invention includes th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/6851C12Q1/701C12Q2531/119C12Q2531/113C12Q2563/107C12Q2547/101C12Q2521/107Y02A50/30
Inventor 王伟伟李潇桐李晓爽孙雪田埂
Owner 北京元码医学检验实验室有限公司