2019-nCoV nucleic acid isothermal amplification primer based on molecular beacon, molecular beacon, test kit and detection method

Abstract

The invention discloses a 2019-nCoV nucleic acid isothermal amplification primer based on a molecular beacon, the molecular beacon, a test kit and a detection method. The 2019-nCoV virus primer comprises an ORF1ab gene primer sequence and an N gene primer sequence of a 2019-nCoV virus; the molecular beacon comprises an ORF1ab gene molecular beacon probe and an N gene molecular beacon probe; the test kit comprises the 2019-nCoV virus primer, a molecular beacon, an NASBA amplification buffer solution and an NASBA enzyme mixed solution, the detection method using the test kit comprises the following steps: firstly, preparing an NASBA amplification buffer solution, preparing an NASBA enzyme mixed solution, preparing a primer mixed solution, and preparing an RNA sample; mixing, reacting and incubating all the solutions, and finally completing detection through a fluorescence detector. The invention shows high-specificity and high-sensitivity detection performance, and a large amount of experiment time is saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer and analytical beacon, a kit and a detection method for rapidly detecting 2019-nCoV virus based on constant temperature amplification technology and molecular beacon technology. Background technique [0002] At present, the more commonly used laboratory diagnostic techniques can be divided into virus cell culture techniques, imaging examinations, electron microscopy examinations, serological examinations, and molecular biological examinations. Among them, virus cell culture isolation is currently the most accurate method, but lacks the sensitivity and speed required for high-throughput screening, which may lead to delays in disease progression; serological methods such as ELISA have low specificity and cannot be diagnosed. Molecular biology detection method is now most common with qRT-PCR method, which has the advantages of high sensitivity, good specificity, wid...

AI Technical Summary

Problems solved by technology

However, due to the relatively high price of the current PCR instrument, this condition is basically available in the laboratories of the top three hospitals, and the technical requirements for operators are relatively high, so it cannot be popularized and used at the grassroots level

In the case of a large-scale outbreak, it is impossible to efficiently screen a large area of ​​people

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