Primer composition for detecting SARS-CoV-2 and application thereof

Abstract

The invention relates to a primer composition for detecting SARS-CoV-2 and application thereof. The primer composition comprises nucleic acid sequences shown as SEQ ID NO: 1 to SEQ ID NO: 12. The primer composition is used for reverse transcription nested PCR; Sanger sequencing is combined; the SARS-CoV-2 gene information can be fast and accurately obtained, so that the fast detection of SARS-CoV-2 and the judgment of SARS-CoV-2 mutant strains can be realized; and good accuracy, sensitivity, specificity and repeatability are realized.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a primer composition for detecting SARS-CoV-2 and an application thereof. Background technique [0002] Coronaviruses belong to the order Nesting Viridae, the family Coronaviridae, and the genus Coronaviridae. They are a class of RNA viruses with an envelope and a linear single-stranded positive strand genome. They are a class of viruses that exist widely in nature. Certain coronaviruses infect humans and cause disease, such as Middle East Respiratory Syndrome (MERS), Severe Acute Respiratory Syndrome (SARS), and pneumonia caused by SARS-CoV-2, which can range from the common cold to severe lung infections. [0003] Pneumonia caused by SARS-CoV-2 is mainly characterized by fever, fatigue and dry cough. A small number of patients have symptoms such as nasal congestion, runny nose, and diarrhea. Detection of SARS-CoV-2 on suspected cases of pneumonia infected by SARS-Co...

AI Technical Summary

Problems solved by technology

[0005] Susceptibility to mutation is an inherent property of most RNA viruses. Reports have revealed the emergence of mutant strains of SARS-CoV-2. SARS-CoV-2 has a proofreading polymerase that can make SARS-CoV -2 has a low probability of base substitution mutations, but SARS-CoV-2 is more prone to deletion mutations, which mostly occur in the recurrent deletion regions (RDRs) of the S protein. As an antibody recognition epitope, deletion mutations of RDRs may lead to changes in the binding specificity of SARS-CoV-2 antibodies, causing antigenic evolution, thereby escaping the neutralizing antibodies induced by pre-mutated SARS-CoV-2

[0006] At present, domestic and foreign detection methods for SARS-CoV-2 mainly focus on two aspects: detection based on PCR amplification of target sequences and sequencing technology (Sanger sequencing, NGS sequencing). The qualitative detection of SARS-CoV-2 nucleic acid is mainly Including reverse transcription + real-time fluorescent quantitative PCR (Real-time PCR), etc., wherein fluorescent PCR technology is simple, fast and sensitive, such as CN111270021A discloses a primer pair, probe, Compositions, kits and applications, the fluorescent RT-RAA primers and fluorescent probes can quickly and qualitatively detect the new coronavirus SARS-CoV-2 in patient samples, but due to the short discovery time of the virus and insufficient scientific knowledge, it is used Gene loci for differential diagnosis have not been validated in large-scale clinical trials, there may be false negatives or false positives, accuracy and analysis-specific risks

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