Detection method of avian influenza virus H5N1

An avian influenza virus and detection method technology, applied in the field of biochemical experiments, can solve the problems of poultry death, infection with human influenza virus, and possible infection with avian influenza virus, etc., and achieves simple operation, low requirements for conditions, and high specificity and sensitivity. Effect

Inactive Publication Date: 2014-03-12
李刚
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  • Abstract
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  • Application Information

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Problems solved by technology

Possibility of mutation One is that two or more viruses enter the same cell for recombination. For example, pigs can infect both human influenza virus and bird influenza virus. Each virus has 8 gene segments. In theory, it can form 256 new recombinant viruses; the second is that the virus gene locus is affected by some factors. In April 1983, low pathogenicity avian influenza in chickens and turkeys caused by the H5N2 virus broke out in Pennsylvania, the United States. Effective control, until October of the same year, the same H5N2 strain suddenly changed from low pathogenicity to high pathogenicity, causing a large number of poultry deaths

Method used

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Embodiment Construction

[0007] The specific steps are as follows: first extract the RNA fragment of the target virus; establish the amplification reaction system: including RNA, DNA polymerization buffer and appropriate reverse transcriptase, etc.; mix the above-mentioned substances with three times the mentioned medical distilled water, and shake slightly; Introduce the primers into it; conduct a warm water bath in an environment of 35 degrees Celsius; perform electrophoresis by agarose gel electrophoresis; high-speed centrifugation; suspend; discard the supernatant; and compare with the HA gene region; The percentage is 1% to 2%. The medical distilled water needs to be warmed at room temperature for at least 24 hours before use, so that its temperature is kept at least 25 degrees Celsius.

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Abstract

The invention relates to a detection method of avian influenza virus H5N1. The detection method is characterized by comprising the following steps of: extracting an RNA (Ribose Nucleic Acid) fragment of target virus; building an amplified reaction system, including RNA and DNA (Desoxvribose Nucleic Acid) polymerized buffer solutions, proper reverse transcriptase, etc.; mixing the substances above with triple extracted medical distilled water; slightly oscillating; introducing a primer; performing warm water bath under a temperature of 35 DEG C; electrically measuring by agarose gel electrophoresis; centrifuging at high speed; suspending; removing supernate; and then comparing with HA (Hyaluronic Acid) gene, wherein the agarose gel electrophoresis is 1-2% in mass percentage; the medical distilled water is warmed for at least 24 hours under room temperature before use so as to keep the temperature in at least 25 DEG C. The detection method has the beneficial effects that the experimental specificity and sensitivity are high; the nucleic acid sequence structure can be described; the operation is relatively simple; and the requirements on conditions are not high.

Description

technical field [0001] The invention relates to a biochemical experiment method, in particular to a method for detecting subtype avian influenza virus H5N1. Background technique [0002] Avian influenza virus (AIV) is a type A influenza virus. Influenza viruses belong to the Orthomyxoviridae family of RNA viruses and are divided into three types: A, B, and C. Among them, influenza A virus is mainly found in poultry, and some subtypes can also infect various mammals such as pigs, horses, seals and whales, as well as humans; influenza B and C viruses are found in seals and pigs, respectively. Generally speaking, there are receptor-specific differences between avian influenza virus and human influenza virus, and avian influenza virus is not easy to infect humans. Individual avian influenza viruses that cause human infection may be mutated viruses. Possibility of mutation One is that two or more viruses enter the same cell for recombination. For example, pigs can infect both...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/70C12Q1/686
Inventor 李刚
Owner 李刚
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