Method for detecting respiratory viruses through next generation sequencing with high flux and application

A second-generation sequencing and detection method technology, which is applied in the field of medical testing, can solve the problems of "false negative" cases and achieve high sensitivity, pollution avoidance and high precision

Pending Publication Date: 2020-05-12
杭州启棣生物技术有限公司
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Problems solved by technology

[0005] In order to solve the problem that real-time quantitative PCR detection will lead to a large number of "false negative" cases, and the need for large sample size detection when epidemics break out, the present invention proposes a high-throughput detection of respiratory viruses using next-generation sequencing. method and the application of this detection method in respiratory virus infection, the detection method has the advantages of high precision, high sensitivity, high throughput and rapid detection

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  • Method for detecting respiratory viruses through next generation sequencing with high flux and application
  • Method for detecting respiratory viruses through next generation sequencing with high flux and application
  • Method for detecting respiratory viruses through next generation sequencing with high flux and application

Examples

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Embodiment 1

[0102] Example 1: Take 2019-nCoV as an example

[0103] (1) Collect specimens and group them into groups, with 96 samples in each group (including positive and negative controls), and record the sample names and other information in an Excel table.

[0104]Nasal swab: Insert one swab into the nose and palate, turn it and take it out carefully, take another swab and take the other nostril in the same way. Dip the swab tip into the sampling tube. Then carry out the pretreatment of nucleic acid extraction: cut the swab, put it into a 2.0ml test tube, add it to 400μl PBS, and take 200μl supernatant sample for extraction after vigorous shaking.

[0105] (2) Extraction of RNA: extract according to the product instructions. Aliquot RNA samples into 96-well plates, 20 μl per well. Then one-step enrichment of nucleic acid target fragments (using SuperScript TM III One-Step RT-PCR System kit as an example):

[0106] (2.1) Set the following program on the PCR instrument.

[0107] ...

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Abstract

The invention relates to the technical field of medical detection, aims to solve the problem that a great number of "false negative" cases can be caused when a real-time quantitative PCR (polymerase chain reaction) method and the requirement that massive samples need to be detected in case of epidemic disease outbreak, and discloses a method for detecting respiratory viruses through next generation sequencing with a high flux and application of the detection method to respiratory virus infection. The detection method comprises the following steps: acquiring samples, and collecting the samplesin test tubes with RNA (ribonucleic acid) protection liquids; extracting RNA by using a RNA extraction kit, and performing RNA reverse transcription and enrichment of nucleic acid in specific areas byusing a one-step method; constructing a DNA (deoxyribonucleic acid) library by using a one-step method, and performing quality testing; performing next generation sequencing; and analyzing sequencesby using a bioinformatic technique. The detection method has the advantages of being high in precision, high in sensitivity, high in flux and rapid in detection.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a method and application for high-throughput detection of respiratory viruses by using next-generation sequencing. Background technique [0002] Respiratory virus infection is a common cause of influenza. Some viruses, such as SARS, MERS, 2019-nCoV, etc., are very harmful to health and the public due to their strong transmission ability, strong pathogenicity, and even high mortality. Big. Different types of influenza have different disease courses, and the prevention and treatment of different types of influenza are also completely different. How to achieve early detection, early diagnosis, early isolation, and early treatment is the focus of epidemic prevention and treatment. [0003] Different influenza has different diagnostic criteria. Taking 2019-nCoV as an example, the current diagnostic criteria are epidemiological history, clinical manifestations (respiratory s...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6869
CPCC12Q1/701C12Q1/6869C12Q2531/113C12Q2535/122
Inventor 董灵胡强
Owner 杭州启棣生物技术有限公司
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