RNA probe for detecting novel coronavirus 2019-nCOV as well as preparation method and application thereof

Abstract

The invention discloses an RNA probe for detecting novel coronavirus 2019-nCOV as well as a preparation method and application thereof, and belongs to the technical field of nucleic acid hybridization. In order to improve the accuracy and specificity of novel coronavirus detection, the invention provides the RNA probe for detecting novel coronavirus 2019-nCOV, and the nucleotide sequence of the RNA probe is shown as SEQ ID No.1. The preparation process comprises the following steps: primer designing, construction of recombinant plasmids, PCR amplification, product purifying, and in-vitro transcription. When the prepared probe is applied to detection of 2019-nCOV, 2019-nCOV virus nucleic acid can be detected at a single cell level, and the detection sensitivity is improved.

Description

technical field [0001] The invention relates to the technical field of nucleic acid hybridization, in particular to an RNA probe for detecting novel coronavirus 2019-nCOV and its preparation method and application. Background technique [0002] The 2019 novel coronavirus (2019-nCoV) has a long incubation period and strong infectivity. It mainly attacks the lungs of animals and causes lung infections. [0003] At present, there are fluorescent quantitative PCR and antibody detection methods for the identification of new coronaviruses. Among them, the nucleic acid detection method is the gold standard for virus detection methods, but there are still many problems in the existing methods, such as: [0004] (1) Fluorescent quantitative PCR can identify nucleic acid in suspected patients. This method requires extraction of RNA in cells, followed by reverse transcription, and then PCR identification. In these experimental operations, the loss of RNA is relatively large. If the sus...

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Problems solved by technology

[0004] (1) Fluorescent quantitative PCR can identify nucleic acid in suspected patients. This method requires extraction of RNA in cells, followed by reverse transcription, and then PCR identification. In these experimental operations, the loss of RNA is relatively large. If the suspected patient is infected In the early stage or the amount of virus contained in the body itself is relatively small, it is easy to cause RNA loss during the extraction process, resulting in false negative results

[0005] (2) Since the new coronavirus is an RNA virus, the mutation speed of RNA virus is fast, and the mutation can continue to accumulate. Fluorescence quantitative PCR experiment has relatively high requirements for nucleic acid sequence, especially in the area covered by PCR primers. If the The nucleic acid in the region is mutated, resulting in a decrease in PCR amplification efficiency or even failure to amplify it, resulting in false negative results

[0006] (3) The fluorescent quantitative PCR experiment has not been sequenced and identified, and non-target fragments may be amplified, resulting in false positives, resulting in inaccurate test results

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