Quick detection technology for H9 type avian influenza virus

An avian influenza virus and detection technology technology, which is applied to the determination/inspection of microorganisms, biological testing, measuring devices, etc., and can solve the problems of long detection period, difficulty in detection, and contamination of tested materials.

Inactive Publication Date: 2009-11-11
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these technical methods have been recognized, adopted and applied, some of them have low sensitivity, some have low specificity, some have low detection rates, some have a long detection period, and some have too much cost, and these Most of the methods need to use live viruses, and there is a hidden danger of spreading the poison to the surrounding environment
Ther

Method used

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  • Quick detection technology for H9 type avian influenza virus
  • Quick detection technology for H9 type avian influenza virus
  • Quick detection technology for H9 type avian influenza virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0026] Embodiment the establishment of H9 type avian influenza virus RT-LAMP method

[0027] 1. Nucleic acid extraction of H9 avian influenza

[0028] Aseptically collect the peripheral blood of poultry infected with H9-type avian influenza virus, add appropriate amount of anticoagulant, add a small amount of pH 7.4, 0.05M phosphate buffer saline (PBS) to the poultry tissue sample, and then use a tissue grinder to homogenize it. Viral RNA was extracted from the processed samples according to the viral RNA extraction kit provided by Treasure Bioengineering (Dalian) Co., Ltd.

[0029] 2. RT-LAMP response to H9 avian influenza

[0030] Two pairs of primers were designed with reference to the sequence of the HA gene of H9 avian influenza registered in GenBank. The present invention adopts the sequence of primer as follows:

[0031] Outer primers:

[0032] Upstream F: 5′-CTACTGTTGGGAGGAAGAGAATGGT-3′

[0033] Downstream B: 5′-GAAAGAATGTGTCCATACCA-3′

[0034] Inside primer:

...

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Abstract

The invention discloses a method for detecting virus from a poultry disease material, and application of the method. The method comprises the following steps: sterilely acquiring tissues of detected dead poultry such as intestinal contents, lung, air chamber, intestines, spleen, liver or heart, or sterilely acquiring blood, trachea swab, cloacae swab or excrement of detected living poultry; extracting RNA from a sample by adopting a Trizol method or a Mini BEST virus RNA extraction kit; using the extracted RNA as a template to carry out an RT-LAMP reaction with upstream and downstream primers at the outside and upstream and downstream primers at the inside, sampling the reactant after the RT-LAMP reaction is finished to carry out agarose gel electrophoresis detection, and judging whether the detected poultry are infected by H9 type avian influenza virus by a way that whether a ladder type band with RT-LAMP reaction specificity appears.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to a method for detecting viruses from poultry disease feed and the application of the method. More precisely, the detection method developed by the present invention using the loop-mediated isothermal amplification technology, from dead poultry disease materials (including intestinal contents, lungs, air sacs, intestines, spleen, liver, heart and other tissues) and live poultry disease materials (including blood, tracheal swabs, cloacal swabs and feces) for the detection of H9 avian influenza virus. Background technique [0002] Avian influenza virus (AIV) has been the most important pathogen affecting the poultry industry. Although the prevalence of avian influenza has been controlled through the widespread use of vaccines, in recent years, H9-type avian influenza that is prevalent in my country has shown atypical symptoms in high-immune chicken flocks, often with a large drop in egg produc...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N27/447G01N33/50
Inventor 刘永生陈豪泰张杰孙德惠马丽娜
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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