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Primer and probe combination and kit for detecting 34 respiratory tract pathogenic microorganisms based on multiple PCR-time-of-flight mass spectrometry

A technology of time-of-flight mass spectrometry and respiratory tract, which is applied to the application field of detection methods in respiratory virus infection, and can solve the problems of high detection cost, long time-consuming, inapplicable triage, and the rapid diagnosis of epidemic prevention and control system.

Pending Publication Date: 2020-11-03
JIANGSU COWIN BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although gene chips and NGS high-throughput sequencing detection technologies have high detection throughput, they are not suitable for the rapid diagnosis of respiratory pathogen monitoring, triage, and epidemic prevention and control systems due to high detection costs and time-consuming. Clinical application

Method used

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  • Primer and probe combination and kit for detecting 34 respiratory tract pathogenic microorganisms based on multiple PCR-time-of-flight mass spectrometry
  • Primer and probe combination and kit for detecting 34 respiratory tract pathogenic microorganisms based on multiple PCR-time-of-flight mass spectrometry
  • Primer and probe combination and kit for detecting 34 respiratory tract pathogenic microorganisms based on multiple PCR-time-of-flight mass spectrometry

Examples

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Embodiment 1

[0094] In the method for highly sensitive detection / or identification of respiratory virus pathogens and novel coronavirus typing methods provided by the embodiments of the present invention, the samples to be tested include positive plasmids corresponding to each site, positive new crown samples and negative sample controls. Including the following steps:

[0095] (1) According to the entire genome sequence of the respiratory virus to be tested, select specific conserved sequences, design PCR primers, and design primers for 34 specific sites. Primers, use a pair of PCR primers, with 10 bases (ACGTTGGATG) sequence at the 5' end of the primers, so that the total length reaches more than 30 bases, in order to distinguish the primers from the probes in terms of molecular weight. Design a single-base extension probe in the conserved sequence region in the amplified region, the length of the probe is 15-29 bases, and at the 3' end of the probe, it is allowed to extend a base determ...

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Abstract

The invention provides a primer and probe combination for multiple respiratory tract pathogen nucleic acid detection and typing identification of new coronaviruses based on multiple PCR time-of-flightmass spectrometry. The primer and probe combination comprises: 1) a primer sequence combination as shown in Table 1; and 2) a probe sequence combination as shown in a table 2. The invention also provides a kit for multiple respiratory tract pathogen nucleic acid detection and typing identification of new coronaviruses based on multiple PCR time-of-flight mass spectrometry. According to the invention, based on a MassARRAY MALDI-TOF MS system, multiple respiratory tract pathogens can be detected at the same time, and typing identification can also be carried out on the 2019 novel coronavirus (2019nCoV), so that the kit has more significance in clinical and virus research.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method for detecting 34 kinds of respiratory pathogenic microorganisms by multiple PCR-time-of-flight mass spectrometry, a matching primer / probe combination and a kit, and the application of the detection method to respiratory virus infection. Background technique [0002] Respiratory tract infections, especially acute respiratory infectious diseases, have always been an important cause of death and disability worldwide, and have received widespread attention worldwide. Viruses are the main cause of respiratory tract infections, and more than 50% of respiratory infections are caused by viruses. Common respiratory infection viruses include influenza A virus, influenza B virus, parainfluenza virus, respiratory syncytial virus, coronavirus, etc. Influenza A virus can infect humans, mammals, and birds. H1N1, H2N2, and H3N2 subtypes mainly infect humans, and other subtypes m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/686C12Q1/6872C12Q1/04C12N15/11C12R1/35C12R1/01
CPCC12Q1/701C12Q1/689C12Q1/686C12Q1/6872C12Q2537/143
Inventor 王冬郭金海王春香
Owner JIANGSU COWIN BIOTECH CO LTD
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