Digital PCR-based novel coronavirus nucleic acid quantitative detection kit and application

Abstract

The invention discloses a digital PCR-based novel coronavirus nucleic acid quantitative detection kit and an application. The reaction total system of the digital PCR-based novel coronavirus nucleic acid quantitative detection kit has 20 ul, comprising 10ul of 2x One-Step RT-ddPCR Supermix, 0.8ul of 25mM manganese acetate solution, 5ul of to-be-detected sample RNA, 1ul of ORFlab gene primer probeworking solution, 1ul of N gene primer probe working solution, 1ul of RPP30 gene primer probe working solution and 1.2ul of Nuclease-Free Water, wherein 5ul of negative control RNA extraction solutionand 5ul of positive control RNA extraction solution are adopted to replace the to-be-detected sample RNA in a negative control reaction system and a positive control reaction system respectively. Based on the innovative RNA one-step reverse transcription microdroplet type digital PCR technology, nucleic acid absolute quantification is carried out specific to highly conservative ORFlab gene and Ngene in the novel coronavirus (2019-nCoV) genome, so that the detection accuracy is improved, and the kit can be used for clinical assisted diagnosis and viral load analysis of novel coronavirus (2019-nCoV) infection, and has a wide clinical application value.

Description

technical field [0001] The invention relates to the technical field of novel coronavirus detection kits, in particular to a digital PCR-based nucleic acid quantitative detection kit for novel coronavirus and its application. Background technique [0002] The new coronavirus was named 2019-nCoV by the WHO, and was named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by the International Committee on Taxonomy of Viruses on February 12, 2020. The virus can cause severe acute respiratory infection (Severe acute respiratory infection, SARI), and its early symptoms are similar to common viral colds. As the disease progresses, patients will experience dyspnea, chest tightness, and even respiratory distress symptoms. Medical imaging examinations show multiple ground-glass opacities in the lungs, and lung consolidation may occur in severe cases. In view of the characteristics of highly contagious and clustered onset of the new coronavirus, timely screening of people in...

AI Technical Summary

Problems solved by technology

Multi-day clinical test results show that this diagnostic method has a high rate of false negatives, and often requires multiple nucleic acid tests before it can be finally verified

At the same time, there may be mutations in this new type of coronavirus (2019-nCoV), and no research on the ORF1ab gene sequence, ORF1ab gene sequence and N gene sequence is involved. The ORF1ab gene is the open reading frame 1ab; the N gene is the nucleocapsid protein; the RPP30 gene is RNase P protein 30KD subunit

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