High throughput detection kit of SARS-CoV-2

Abstract

The invention discloses a high throughput detection kit of SARS-CoV-2. The kit comprises 274 primers, wherein nucleotide sequences of the 274 primers are successively shown by SEQ ID NO: 1 to SEQ ID NO: 274. According to the invention, a multiple PCR technology and an NGS sequencing technology are used for million-multiple enrichment of SARS-CoV-2 analysis; through setting of external parameters and internal parameters of a known copy number, whether the SARS-CoV-2 exists or not can be detected sensitively, and a concentration of the SARS-CoV-2 can be judged; and total length data of the SARS-CoV-2 can be obtained, and research on evolution and variation of the SARS-CoV-2 is of high sensitivity, specificity and accuracy. Meanwhile, reverse transcription and the multiple PCR technology arecombined into one step, so that risks in SARS-CoV-2RNA degradation are reduced. The detection kit has important application values.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a high-throughput detection kit for SARS-CoV-2. Background technique [0002] SARS-CoV-2 belongs to betacoronavirus, and its transmission ability is stronger than that of SARS. Early detection and rapid detection of SARS-CoV-2 are not only very important for the diagnosis of patients, but also play a key role in effectively preventing the further spread of the epidemic, especially in the detection of asymptomatic or latent virus carriers. [0003] At present, the routine detection method of SARS-CoV-2 is real-time fluorescent RT-PCR detection, and its nucleic acid detection is mainly aimed at the open reading frame La / b (Openred frame Lab, ORF Lab) and nuleocapsid protein (nuleocapsid protein) in the virus genome. , N). RT-PCR will take one or several of the targets to detect the virus, which plays a pivotal role in the confirmed patients, but because the βcoronavirus is a single-s...

AI Technical Summary

Problems solved by technology

The sequencing method based on the whole genome can obtain all the nucleic acid data in the sample, but the disadvantage is that the exogenous nucleic acid data account for too much, and the required target virus (SARS-CoV-2) data that can be obtained is small, especially in the case of weak positive , it is easy to miss the detection

The whole genome sequencing method based on probe capture can first enrich the target virus (SARS-CoV-2) tens of thousands of times, and then perform sequencing, but the library construction process is very cumbersome and takes a long time, which is not conducive to the virus prevention and control

The whole genome capture method based on multiplex PCR uses specific primers to enrich the target virus by a million times, and then performs sequencing to obtain the whole genome data of the virus. It is easy to operate, has very high sensitivity, and can capture very weak positive The sample is detected, but it cannot judge the positive degree of the sample, that is, the relative concentration of the virus

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