A kind of specific primer and application for novel coronavirus detection

Abstract

The invention discloses a specific primer for the detection of novel coronavirus and its application, comprising the steps of: inactivating the collected samples, and extracting viral RNA; reversing the qualified viral RNA extracted through the step S1 and performing cDNA synthesis ; Use specific primers to perform fragment amplification to form a PCR reaction product sample; mark the PCR reaction product sample, and construct a sequencing library; perform nanopore sequencing operations on the sequencing library; the sequencing quality of the present invention is good, and the obtained When the sample is compared to the new crown genome, a reads comparison rate of up to 94.16% can be achieved, which can assist in determining the infection status of the patient and achieve rapid and accurate detection of the new coronavirus.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a specific primer for detection of novel coronavirus and its application. Background technique [0002] The novel coronavirus (2019-nCoV) infection is currently highly transmissible and has a high fatality rate, which is a major public health issue of national concern. At the same time, it is also the epidemic season of other respiratory viruses, which has a huge impact on clinical diagnosis and differential diagnosis. [0003] In view of the high prevalence, distribution and widespread nature of coronaviruses, the diversity of genes and frequent recombination of genomes, and the increasing interaction between humans and animals, it is very likely that new coronaviruses will appear periodically in humans, Therefore, there is an urgent need to study methods for its rapid detection. [0004] The most rapid and effective nucleic acid detection method in the prior art is RT-PCR,...

AI Technical Summary

Problems solved by technology

[0004] The most rapid and effective nucleic acid detection method in the prior art is RT-PCR, and the specific technology used is the new coronavirus RT-PCR kit, but this method has certain defects. According to the RT-PCR system provided by the kit manual Add one to three pairs of primers with different specificity to check the specificity, the detection area is ORF1ab, N area and E area

Since the entire genome region of 2019-nCoV is not completely covered, its accuracy is affected to a certain extent, and RNA viruses are prone to mutation and recombination. At present, there have been many cases of positive patients with negative RT-PCR tests.

For differential diagnosis or a large number of samples to be tested, a large amount of reagents, consumables and time will be consumed when there are many testing items. In addition, the resulting missed screening will also bring corresponding difficulties to subsequent diagnosis and prevention and control.

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