Reaction system, method and application for detecting low-abundance novel coronavirus
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, control, etc., achieve stable detection rate, reduce crossover Contamination probability and effect of shortening detection time
Active Publication Date: 2020-11-03
北京元码医学检验实验室有限公司
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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
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[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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The invention discloses a reaction system, method and application for detecting low-abundance novel coronaviruses. The invention enables the isothermal amplification reaction and the PCR amplification reaction to be carried out in the same reaction system, avoids the mutual influence between the two, and realizes the purpose of detecting low-abundance nucleic acid. It is more than an order of magnitude lower than the lower limit of the single PCR detection method, which further increases the sensitivity and specificity of the detection and stabilizes the detection rate. At the same time, the quantitative analysis of the viral load is carried out through the CT value of the qPCR amplification curve. By integrating the specific signal amplification of two sets of reactions and the one-tube design of high-sensitivity fluorescence detection, the detection time is shortened to within half an hour, which is more than 3 times shorter than conventional detection, so as to achieve the purpose of rapid and stable detection of extremely low-abundance pathogens. Prevent the missed detection problem in the detection of trace pathogenic samples.
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 the foll...
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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