2019-nCoV nucleic acid isothermal amplification detection kit based on SYBR Green I and detection method

Abstract

The invention discloses a 2019-nCoV nucleic acid isothermal amplification detection kit based on SYBR Green I and a detection method. The kit comprises an ORF1ab gene primer sequence and an N gene primer sequence of a 2019-nCoV virus, an SYBR Green I dye, an NASBA amplification buffer solution and an NASBA enzyme mixed solution. The detection method using the kit comprises the following steps: firstly, preparing the NASBA amplification buffer solution, preparing the NASBA enzyme mixed solution, preparing a primer mixed solution and preparing an RNA sample; and carrying out mixed reaction on all the solutions, carrying out incubating, and finally completing detection by using a fluorescence detector. The kit and the method show high-specificity and high-sensitivity detection performance, and a large amount of experimental time is saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection kit and a detection method for rapid detection of 2019-nCoV virus based on constant temperature amplification technology and SYBR Green I 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, wide linear range and accura...

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 tertiary hospitals, and the technical requirements for operators are relatively high, so it cannot be popularized and used at the grassroots level

When a large-scale outbreak occurs, it is impossible to efficiently screen a large number of people

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