Method, primer group and kit for capturing novel coronavirus whole genome

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] In late December 2019, a patient with viral pneumonia caused by an unknown microbial pathogen was reported in Wuhan, China, and a novel coronavirus was subsequently identified as the causative pathogen. The study showed that the 10 genome sequences of the novel coronavirus initially obtained from 9 patients were extremely similar, with a sequence identity of more than 99.98%. It is worth noting that this novel coronavirus is closely related (88% concordance) to two SARS-like coronaviruses, bat-SL-CoVZC45 and bat-SL-CoVZXC21, collected in Zhoushan, eastern China in 2018, but not closely related to SARS. - CoV (about 79%) and MERS-CoV (about 50%) are more distant. This novel coronavirus is sufficiently different from SARS-CoV to be considered a novel human-infecting oronas...

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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