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Multiple RT-qPCR kit and primer probe composition for coronavirus typing detection and use method of RT-qPCR kit and primer probe composition

A coronavirus, primer and probe technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., can solve problems such as death, achieve high sensitivity, good typing effect, and short detection cycle Effect

Pending Publication Date: 2020-08-11
NINGBO HEALTH GENE TECHNOLOGIES CO LTD +2
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  • Abstract
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Problems solved by technology

[0002] Common signs of human infection with the novel coronavirus (SARS-CoV-2) include respiratory symptoms, fever, cough, shortness of breath, and dyspnea, and in severe cases, the infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death , so far there is no specific treatment, early detection and early isolation is the most effective measure to curb the spread of the virus

Method used

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  • Multiple RT-qPCR kit and primer probe composition for coronavirus typing detection and use method of RT-qPCR kit and primer probe composition
  • Multiple RT-qPCR kit and primer probe composition for coronavirus typing detection and use method of RT-qPCR kit and primer probe composition
  • Multiple RT-qPCR kit and primer probe composition for coronavirus typing detection and use method of RT-qPCR kit and primer probe composition

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Embodiment 1

[0043] The coronavirus multiplex RT-qPCR genotyping kit was used to detect SARS-CoV-2 positive confirmed patients (sample number S1), HCoV-229E positive confirmed patients (sample number S2), HCoV-NL63 positive confirmed patients (sample number S3 ), HCoV-HKU1-positive confirmed patients (sample number S4), HCoV-OC43-positive confirmed patients (sample number S5) and non-suspected patients (sample number S6) in the presence of coronavirus RNA in respiratory samples:

[0044] 1. Nucleic acid extraction: Use nucleic acid extraction reagents to automatically extract nucleic acids from samples S1 to S6 on an automatic nucleic acid extraction instrument at the same time, and store the extracted nucleic acid samples at -20°C;

[0045] 2. Prepare the reaction system according to Table 5, perform RT-qPCR according to Table 6, and obtain the fluorescence amplification curve;

[0046] 3. Determine the type of coronavirus contained in the sample according to the judgment standard.

[00...

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Abstract

The invention discloses a multiple reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) kit and a special primer probe composition for distinguishing a novel coronavirus (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV) and four kinds of common human coronaviruses (including human coronavirus 229E (HCoV-229E), human coronavirus NL63 (HCoV-NL63), human coronavirus HKU1 (HCoV-HKU1) and human coronavirus OC43 (HCoV-OC43), and a use method of the kit and the composition. According to the kit, a single-tube four-color fluorescence multiple RT-qPCR method is adopted for detecting and distinguishing coronaviruses aiming at a human respiratory tract sample. The coronavirus typing detection kit disclosed by the invention is short in detection period, high insensitivity and good in typing effect; quality monitoring of sample extraction and amplification processes can be realized by monitoring human RNA internal reference B2M genes, and false positive results caused by aerosol pollution are reduced by utilizing a dUTP-UNG enzyme anti-pollution system.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection, in particular to a multiplex RT-qPCR kit for typing and detecting coronaviruses, a primer-probe composition and a method for using the same. Background technique [0002] Common signs of human infection with the novel coronavirus (SARS-CoV-2) include respiratory symptoms, fever, cough, shortness of breath, and dyspnea, and in severe cases, the infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death , so far there is no specific treatment, early detection and early isolation is the most effective measure to curb the spread of the virus. Due to the essential differences in the diagnosis and treatment methods for patients with different coronavirus infections, distinguishing common coronavirus infections and performing shunt treatment will help concentrate medical resources for targeted treatment of SARS-CoV-2 positive patients and help impro...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2600/166C12Q2537/143C12Q2521/107C12Q2521/531C12Q2545/113C12Q2545/101C12Q2563/107Y02A50/30
Inventor 冯志山李贵霞张迪骏王乐赵梦川郭映辉帖彦清段素霞吴勇
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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