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Method for detecting influenza virus by liquid phase chip

A liquid phase chip detection technology for influenza virus, applied in the field of virus detection, can solve the problems of lack of sensitivity, failure to rule out influenza virus in patients, and inability to identify genetic material, etc., to reduce non-specific annealing of primers, fast detection speed, high The effect of specificity and sensitivity

Active Publication Date: 2010-06-02
SHANDONG ACV BIOTECH CO LTD
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AI Technical Summary

Problems solved by technology

Existing PCR tests have good specificity but lack sensitivity because virus may not be present in patient specimens or genetic material cannot be identified due to failure of amplification or detection
And a negative test does not rule out the possibility that a patient has the flu virus
And they don't allow rapid and accurate typing of influenza viruses

Method used

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  • Method for detecting influenza virus by liquid phase chip
  • Method for detecting influenza virus by liquid phase chip
  • Method for detecting influenza virus by liquid phase chip

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Experimental program
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Effect test

Embodiment Construction

[0020] 1. Synthesis of primers and probes

[0021] Using the knowledge of bioinformatics and related bioinformatics software, the non-structural nucleic acid sequences of all influenza A, influenza B and H1, H2, H3, H5, H7 of influenza A that can be retrieved in Genbank are analyzed. , H9 and N1, N2 subtype nucleic acid sequence homology analysis, find out the conserved regions, design primers and specific probes for the corresponding gene fragments.

[0022] The primers use improved nested primers, that is, a shorter outer primer (15-20bp) and a longer inner primer (35-50bp) with a universal primer (15-20bp) at the 5' end. One 5' end is labeled with biotin. The 5' end of the specific probe is aminated and coupled to the fluorescently encoded microspheres.

[0023] The final synthesized primer and probe sequences are as follows:

[0024] Outer primer:

[0025] H1: 5'-AGG GCA TTT CGC TGA CTA TGA

[0026] 5’-ATT TAT GCT TGG CGG GTG AT

[0027] H2: 5'-TGA TAG GCT TCT AA...

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Abstract

The invention discloses a method for detecting influenza virus by a liquid phase chip. The method is used for carrying out homology analysis on all Non Structural nucleotide sequences of A type influenza virus and B type influenza virus as well as H1, H2, H3, H5, H7 and H9 of the A type influenza virus and hypotypes of N1 and N2, and primers and specific probes are designed aiming at the corresponding gene segments; an improved nest-PCR method is adopted for PCR amplification by using nested primers with different lengths; amination treatment is carried out the 5' end of the specific probe, and the specific probe is coupled with a fluorescent coded microballoon; incubation hybridization is carried out on the PCR amplification product and the mixed microballoon coupled with the specific probe; and finally Luminex 100 is used for detecting. The detection method has higher specificity and sensitivity as well as rapid detection speed, and can be used for indentifying and monitoring the influenza viruses.

Description

(1) Technical field [0001] The present invention relates to a method for detecting virus, in particular to a method for detecting influenza virus by using a liquid phase chip. (2) Background technology [0002] Influenza (flu) is an acute respiratory infectious disease caused by influenza virus. Since 2005, cases of poultry and human infection with influenza virus have occurred all over the world. After SARS, bird flu caused a huge panic to people all over the world. A key issue in controlling influenza epidemics is timely and accurate differential diagnosis, which requires very accurate and sensitive diagnostic methods. [0003] Influenza virus belongs to Orthomyxovirus. According to the difference of the soluble complement-binding antigen of nucleoprotein, influenza virus can be divided into three types: A (A), B (B), and C (C). Type A influenza virus is further divided into several subtypes according to the antigenicity of surface hemagglutinin (Hemagglutinin HA) and ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64C12R1/93
Inventor 汪朝晖王华林张凯宁张国宁李艳艳
Owner SHANDONG ACV BIOTECH CO LTD
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