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Probe and primer composition for simultaneously detecting human novel coronavirus, influenza A virus and influenza B virus

A technology of influenza B virus and primer composition, applied in the field of in vitro diagnosis and biological detection, can solve the problems of low practicability, low sensitivity, and complicated methods of clinical detection, and achieve suitable for popularization and application, high reference value, The effect of high sensitivity

Pending Publication Date: 2020-07-14
JIANGSU BIOPERFECTUS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional detection of influenza virus and human coronavirus mainly includes serological detection, antigen detection, virus isolation and electron microscopy detection, etc., but the above methods are not practical for clinical detection due to the shortcomings of time-consuming detection, complicated methods, poor specificity, and low sensitivity. high
At present, there is a lack of kits for the detection of probes and primer compositions for the simultaneous detection of human novel coronavirus and influenza A and influenza B viruses on the market.

Method used

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  • Probe and primer composition for simultaneously detecting human novel coronavirus, influenza A virus and influenza B virus
  • Probe and primer composition for simultaneously detecting human novel coronavirus, influenza A virus and influenza B virus
  • Probe and primer composition for simultaneously detecting human novel coronavirus, influenza A virus and influenza B virus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: the design of primer, probe:

[0040] Look up the M gene sequence of influenza A virus, the NS gene sequence of influenza B virus, and the genome sequence of RNaseP gene through NCBI, perform sequence comparison through Primer Express3.0.1, design primers and probes with BioEdit, and confirm primer probes with Blast Specificity, get primers and probes for Influenza A, Influenza B, RNaseP; designed on the basis of primers and probes for ORF 1ab and N gene of novel coronavirus (2019-nCoV) announced by the National Center for Disease Control and Prevention Primers and probes for ORF1ab and N genes.

[0041] Specific primers and probes for rapid identification of influenza A virus, influenza B virus and novel coronavirus (2019-nCoV), including:

[0042] (1) Primer probes for amplifying influenza A virus, preferred primer probe concentrations are respectively 200nM, 200nM, 100nM, and the sequences are as follows:

[0043] Primer SEQ1: 5'-GAAACGTAYGTTTCTCTCTAT...

Embodiment 2

[0062] Embodiment 2: the extraction of template RNA

[0063] (1) Collection of samples, including throat swabs, nasal swabs, nasopharyngeal extracts, sputum, respiratory tract extracts, bronchial lavage fluid, alveolar lavage fluid, lung tissue biopsy specimens, blood samples, serum samples, collection Methods as below:

[0064] ① Pharyngeal swab: Use 2 plastic rod swabs with polypropylene fiber heads to wipe the bilateral pharyngeal tonsils and the posterior pharyngeal wall at the same time, and immerse the swab heads in 3ml of virus preservation solution (isotonic saline solution, tissue culture solution can also be used) or phosphate buffer), discard the tail, and screw the cap tightly.

[0065] ② Nasal swab: Gently insert a plastic rod swab with a polypropylene fiber head into the nasal palate in the nasal passage, stay for a while, and then slowly turn it out. Take another plastic rod swab with polypropylene fiber head and collect the other nostril in the same way. The...

Embodiment 3

[0073] Embodiment 3: the preparation of primer probe reaction solution and kit reaction solution

[0074] (1) Preparation of primer probe reaction solution:

[0075]

[0076] (2) Preparation of kit reaction solution:

[0077]

[0078]

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PUM

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Abstract

The invention discloses a probe and primer composition for simultaneously detecting human novel coronavirus, influenza A virus and influenza B virus. 10 specific primers and 5 probes are designed mainly according to genome specificity of M gene of influenza A virus, NS gene of influenza B virus, ORF1ab gene, N genes and internal standard gene RNaseP of novel coronavirus (2019-nCoV), and influenzaA virus, influenza B virus and novel coronavirus 2019-nCoV are distinguished according to amplification curves of different channels by utilizing a real-time fluorescence PCR amplification technology.

Description

Technical field: [0001] The invention belongs to the technical field of in vitro diagnosis, specifically relates to the technical field of biological detection, in particular to a probe and primer composition for simultaneously detecting human novel coronavirus, influenza A and influenza B viruses. Background technique: [0002] Influenza virus belongs to the Orthomyxoviridae family (Orthomyxoviridae), referred to as influenza virus, including human influenza virus and animal influenza virus, human influenza virus is divided into A (A), B (B), C (C) Type, is the causative agent of influenza (flu). Among them, influenza A virus can naturally infect humans and various animals, often occurs suddenly and spreads rapidly, and has caused worldwide pandemics many times; influenza B virus only infects humans under natural conditions, and is generally sporadic, outbreak or small epidemic; Influenza C virus often infects children, mostly sporadic, sporadic or outbreaks, but not popul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2563/107C12Q2561/113
Inventor 曹新文杨静李秀林王政峰赵西浩崔东明李青范东颖张蓉刘中华王国强
Owner JIANGSU BIOPERFECTUS TECH CO LTD
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