COVID-19 (coronavirus disease 2019) nucleic acid testing method capable of increasing accurate rate

A technology for COVID-19 and coronavirus, applied in the direction of biochemical equipment and methods, measurement/testing of microorganisms, and resistance to vector-borne diseases, etc., can solve the problems of infection in patients with polycolloidal gold method, and achieve reduced detection, The effect of improving sensitivity and expanding sensitivity

Pending Publication Date: 2020-04-28
南京申基医药科技有限公司
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Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies in the prior art, including the problems that there are many false negatives in the existing fluorescence quantitative PCR method nucleic acid detection and the colloidal gold method detection cannot accurately predict the patient's inf

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  • COVID-19 (coronavirus disease 2019) nucleic acid testing method capable of increasing accurate rate
  • COVID-19 (coronavirus disease 2019) nucleic acid testing method capable of increasing accurate rate
  • COVID-19 (coronavirus disease 2019) nucleic acid testing method capable of increasing accurate rate

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Embodiment Construction

[0040] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present invention, but not all of them.

[0041] A COVID-19 novel coronavirus nucleic acid detection method capable of improving accuracy, specifically comprising the following steps: Step S1: Mixing an appropriate amount of nucleic acid extract as a sample to be tested with a reverse reaction system for reverse reaction, the reverse reaction system including RNA reverse transcription Enzymes, recombinases capable of binding single-stranded nucleic acids, single-stranded DNA-binding proteins (SSB) and strand-displacing DNA polymerases, allowing RNA with 3' terminal polyA to be reverse-transcribed with the help of reverse transcriptase and oligo (dT) cDNA of poly(T); Step S2: Perform RPA reaction on the reversed cDNA for a period of time to further amplify, use the protein-DNA compl...

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Abstract

The invention relates to a COVID-19 (coronavirus disease 2019) nucleic acid testing method capable of increasing the accurate rate. firstly, a reversal reaction is performed in a reversal reaction system, so that RNA of polyA is subjected to reverse transcription under the assistance of reverse transcriptase and oligo (dT) to form cDNA of poly (T); meanwhile, cDNA subjected to reverse transcription is subjected to an RPA reaction for a period of time for further amplification; an amplified product is mixed with a QPCR (real-time quantitative PCR detecting system), and the mixture is subjectedto a chain reaction on a QPCR instrument and then sequentially subjected to denaturation, reannealing and final extended chain circular reaction respectively; fluorescence signal intensity is collected and detected through detection equipment, qualitative or semiquantitative analytic determination on target segments, and finally, the virus detection result of a patient is judged according to obtained different fluorescence signal data. According to the technical scheme, the COVID-19 nucleic acid testing method which is high in specificity, accurate is result, low in false negative probability,capable of monitoring each stage from the early period to the later period of infection of the patient and capable of increasing the accurate rate can be provided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid detection method for COVID-19 novel coronavirus that can improve accuracy. Background technique [0002] Novel coronavirus pneumonia (NCP for short) is an acute respiratory infectious disease caused by 2019 novel coronavirus (COVID-19) infection. About half of the patients developed dyspnea more than a week later, and severe cases rapidly progressed to acute respiratory distress syndrome, septic shock, metabolic acidosis that was difficult to correct, and coagulation dysfunction. Since the transmission route of the new coronavirus includes direct transmission, aerosol transmission and contact transmission, it is highly contagious, coupled with its long incubation period, it has brought great challenges to the epidemic prevention work. [0003] NCP is a single-stranded RNA virus. It does not have the ribosome system required for RNA virus translation. After entering th...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844
CPCC12Q1/701C12Q1/6844C12Q2521/107C12Q2521/507C12Q2522/101C12Q2563/107C12Q2531/119C12Q2537/1376Y02A50/30
Inventor 童坤乔森肖潇黄磊张硕
Owner 南京申基医药科技有限公司
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