High-sensitivity novel coronavirus 2019-nCoV nucleic acid detection kit and use method

Abstract

The invention discloses a high-sensitivity novel coronavirus 2019-nCoV nucleic acid detection kit and a use method. The high-sensitivity novel coronavirus 2019-nCoV nucleic acid detection kit comprises an ORF1ab gene primer pair, an N gene primer pair, an endogenous reference gene primer pair and a probe set. Novel coronavirus (2019-nCoV) ORF1ab and N genes are used as target areas, specific primers and probes are designed; by detecting the change of fluorescence signals, the novel coronavirus 2019-nCoV nucleic acid in a sample is subjected to qualitative detection; the 5' ends of the ORF1ab gene probe and the N gene probe are modified by a fluorophore FAM to form a monochromatic double-target detection system, and a large-volume system higher than the conventional system is used for amplification, so that when nucleic acid detection is carried out on the novel coronavirus 2019-nCoV, the detection sensitivity is high and can reach 50 copies/mL; and moreover, the nucleic acid extraction and detection process is monitored by taking a ribonuclease P (RNase P, RNP) gene widely existing in normal human epithelial cells as an internal target gene, and false negative and false positive are avoided, so that the nucleic acid detection kit is wide in market popularization prospect.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a high-sensitivity novel coronavirus 2019-nCoV nucleic acid detection kit and its use method. Background technique [0002] The main route of transmission of 2019-nCoV is through respiratory droplets, excreted in respiratory secretions, infected through oral fluid, sneezing, and can also be transmitted through contact. The incubation period after infection with the new coronavirus is 3-7 days, generally no more than 14 days. The clinical manifestations are mainly fever, fatigue, dry cough, and upper respiratory symptoms such as nasal congestion and runny nose are rare. About half of the patients have dyspnea after more than a week, and severe cases progress Acute respiratory distress syndrome, shock, uncorrectable metabolic acidosis, and coagulopathy. [0003] Coronavirus is a single-stranded RNA virus, belonging to Nidovirales (Nidovirales) Coronaviridae (Coronavir...

AI Technical Summary

Problems solved by technology

However, the above methods detect different target genes separately, and the detection system of the existing method is small, the sample amount of the template is small, and the detection sensitivity is insufficient, so there is still the problem of missed detection

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