Pseudo-virus standard substance for nucleic acid diagnosis of novel coronavirus 2019-nCov and application thereof

Abstract

The invention relates to a pseudo-virus standard substance for nucleic acid diagnosis of novel coronavirus 2019-nCov and application thereof. The pseudo-virus standard substance is prepared through the steps of gene synthesis, plasmid extraction, pseudo-virus packaging and the like. Wherein the synthesized gene comprises a novel coronavirus 2019-nCov gene RdRp, a Gene E and a Gene N; the 2019-nCovpseudo-virus standard substance is provided for the first time, is different from RNA synthesized in vitro and clinical positive living virus in the past, perfectly overcomes the defects of RNA and clinical positive living virus, is really suitable for performance evaluation of test kit, comprises limit of detection, specificity, repeatability and the like, and is further applied to clinical diagnosis of 2019-nCov; the pseudo-virus standard substance has the advantages of no pathogenicity, reproducibility, reliable quality control method and stable batch-to-batch, can be stably prepared and supplied for a long time, requires the biological safety level of P2 in a laboratory, and meets the safety requirements of many units.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a pseudovirus standard for nucleic acid diagnosis of novel coronavirus 2019-nCov and its application. Background technique [0002] The new type of coronavirus is named 2019-nCov by the World Health Organization, which can cause severe acute respiratory infection (Severe acute respiratory infection, SARI). Its early symptoms are similar to common viral colds. Distress symptoms, multiple ground-glass opacities in the lungs can be seen through medical imaging examinations, and lung consolidation may occur in severe cases. 2019-nCov, as a new type of coronavirus, is a type of RNA virus with an envelope and a linear single-stranded positive strand genome. The relevant sequences of the virus samples collected from patients in different regions have also been released to the 2019-nCoV information database (https: / / bigd.big.ac.cn / ncov). [0003] The sequence included in NCBI is NC...

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Problems solved by technology

However, both of them have obvious certainties. First, the RNA synthesized in vitro is generally very pure, single, does not involve extraction, and does not conform to the microenvironment of clinical viruses. Therefore, the detection sensitivity will inevitably be overestimated, which is also a high probability of false detection. The main reason for negative; clinically positive virus again, even if it is an inactivated virus, there is a great biological safety hazard, so the laboratory level is required to be P3-P4, and most of the development of diagnostic kits do not meet this requirement. In addition The source of clinically positive virus is also very limited and cannot be supplied stably, so it is repeatedly used for the performance evaluation of the kit

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