A kind of probe and primer set, test kit, detection method utilizing L/RPA rapid detection sars-cov-2

Abstract

The invention relates to the field of biological detection, in particular to a probe, a primer set, a kit and a detection method for rapidly detecting SARS-CoV-2 by using L / RPA. A probe and primer set that utilize L / RPA to rapidly detect SARS-CoV-2 disclosed by the present invention are respectively a probe and a primer set for the N gene and an ORF1ab gene; the probe and the primer set for the N gene Including: probe set: N-Probe2A-3, N-Probe2B-3, primer set N-Primer3F, N-Primer3R; the probe and primer set for ORF1ab gene include: probe set: O-Probe1A-2 , O-Probe1B, primer sets O-Primer2F, O-Primer2R, the present invention also discloses a detection method that can achieve sensitivity equivalent to that of RT-qPCR without a reverse transcription step within 30min. It is fast and sensitive, easy to operate, and avoids sample contamination, and is more expected to be applied to the detection of virus mutant strains.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a probe, a primer set, a kit and a detection method for rapidly detecting SARS-CoV-2 by using L / RPA. Background technique [0002] The novel coronavirus (SARS-CoV-2) has posed a serious threat to the health and safety of people all over the world. Effective vaccines, accurate diagnostics are critical to controlling infectious diseases. Under the premise of in-depth genomics research at this stage, the molecular diagnostic technology for the new coronavirus has also achieved rapid development. At present, the most important detection method for clinical molecular diagnosis is reverse transcription fluorescent quantitative PCR method (RT-qPCR), which transcribes coronavirus RNA into cDNA, and performs specific detection of coronavirus biomarkers through polymerase chain reaction. Amplification, and finally real-time detection of fluorescent signals. However, the RT-qPCR detec...

AI Technical Summary

Problems solved by technology

[0003] However, molecular diagnostic methods developed based on PCR, LAMP, and RPA technologies can only amplify DNA templates, but cannot directly amplify RNA templates. Therefore, viral RNA must be reverse-transcribed into cDNA for subsequent amplification.

As we all know, RNA is easily polluted by RNase in the environment, and the reverse transcription experiment itself has high requirements for the concentration and purity of RNA, which directly requires operators not only to be professional enough, but also to be careful enough to avoid environmental treatment Therefore, the application of molecular detection methods based on the reverse transcription step is greatly limited.

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