A novel coronavirus whole genome capture method, primer set and kit

Abstract

The invention relates to a super-sensitivity method, primer group and kit for capturing whole genome of novel coronavirus. The method can conveniently and quickly amplify the whole genome of the novelcoronavirus by using a small amount of RNA, and can directly dock with a second-generation sequencing library-building reagent and a third-generation sequencing platform so as to obtain a whole genome sequence of the novel coronavirus.

Description

technical field [0001] The invention belongs to the field of virus detection, and in particular relates to a novel coronavirus whole genome capture method, primer set and kit. Background technique [0002] Coronaviruses are non-segmented single-stranded positive-sense RNA viruses ranging in length from 26,000 to 32,000 bases. According to the serotype and genome characteristics, the subfamily Coronaviridae is divided into α, β, γ, and δ four belongs to. SARS-CoV-2 belongs to the genus β, with a full length of 29847bp. [0003] Since January 26, 2020, seven new coronavirus nucleic acid detection kit products have successively passed the emergency approval of the State Drug Administration and obtained marketing approval. The SARS-CoV-2 identification kit is mainly based on the fluorescent quantitative PCR method. Practice has shown that it has high detection specificity, low cost, and easy operation. However, this type of kit designs specific primers and probes based on kno...

AI Technical Summary

Problems solved by technology

However, the differences in application scenarios and many complex factors jointly affect the success rate of obtaining the whole genome of pathogens. At the same time, there are also problems such as time, manpower, and high sequencing costs for front-line personnel under the heavy task of testing.

Therefore, pathogenic metagenomic sequencing (mNGS) can be used for the diagnosis of negative samples, but it is not easy to obtain the whole genome information, and there are reasons such as complicated operation and relatively long detection time (requiring 24-72 hours). It is impossible to achieve the purpose of large-scale promotion and rapid diagnosis

As a technical supplement, the genome capture method based on amplification is a "Culture-free" virus genome-specific enrichment sequencing solution, which is easy to operate and low in cost, and is suitable for the current needs of epidemic prevention and control. However, existing The operation steps of the probe capture sequencing method are complicated, mainly including 7 different technical operations, and each part of the operation requires more than 5 steps, and many can even reach 33 steps. Once the operation is performed, in order to meet the detection requirements, Operations within each section cannot be paused and must be completed continuously

This brings a lot of uncontrollability and operational complexity to the actual operation.

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