Polymerase chain reaction PCR detection method of largemouth black bass ulcer syndrome virus

A syndrome virus and chain reaction technology, which is applied in the field of specific detection of largemouth bass ulcer syndrome iridescent virus, can solve the problem of inability to distinguish largemouth bass ulcer syndrome virus, etc., and achieves fast detection speed and detection accuracy. High, low cost effect

Inactive Publication Date: 2010-09-22
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

But the method was not yet able to distinguish largemouth bass ulcer syndrome virus from other viruses in the Ranavirus genus

Method used

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Embodiment Construction

[0029] The specific implementation steps of the largemouth bass ulcer syndrome virus polymerase chain reaction PCR detection method are as follows:

[0030] (1) Design of primers

[0031] Using bioinformatics methods, the major capsid protein (MCP) gene sequence of largemouth bass ulcer syndrome virus was compared with the known iridescent virus MCP gene sequence, and a pair of specifically amplified largemouth bass ulcer virus MCP genes were designed. The primer of the fragment, the gene sequence of the primer is:

[0032] P1: 5'-TCTGTTACGGGTTCTGGCATC-3'

[0033] P2: 5'-CCAGCCAAGAGTTGAGCACAT-3'

[0034] A 241bp fragment was expected to be amplified using the viral genome as a template. The homology of the primer sequence with the corresponding sequence of the epidemic hematopoietic organ necrosis virus MCP (the closest relative to the largemouth bass ulcer syndrome virus reported in China) is only 69%, so the primer is considered to be a specific primer.

[0035] In the a...

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Abstract

The invention provides a polymerase chain reaction PCR detection method of a largemouth black bass ulcer syndrome virus, comprising the following steps of: comparing the MPC (Main Capsid Protein) gene sequence of the largemouth black bass ulcer virus with the MPC gene sequence of an unknown iridovirus, and designing and synthetizing the following primers: P1: 5'-TCTGTTACGGGTTCTGGCATC-3' and P2: 5'-CCAGCCAAGAGTTGAGCACAT-3'; amplifying a segment 241bp; and carrying out template DNA (deoxyribonucleic acid) extraction, PCR reaction and agarose gel electrophoresis detection, and detecting that the lowest density of plasmids is 10[4] copy number / muL after the PCR reaction is carried out for 30 rounds. The method has the technical characteristics of rapid detection, high precision and low cost and is suitable for being popularized and applied in a wide range.

Description

technical field [0001] The invention relates to a specific detection method for largemouth bass ulcer syndrome iridescent virus. Background technique [0002] Largemouth bass (Micropterus salmoides), also known as California bass, has the characteristics of fast growth, less disease, low temperature resistance, delicious meat and easy to catch. It has been promoted to all parts of the world since the 1970s. The production is around 100,000 tons. However, from June to October every year since 2006, a kind of ulcer syndrome has been prevalent in the cultured largemouth bass in Guangdong Province, and it has a tendency of getting worse year by year. The skin and muscles of fish infected with the virus are festered, the spleen and kidney become enlarged, and the mortality rate of diseased fish is as high as 60%, which has caused huge losses to farmers and has posed a serious threat to the largemouth bass farming industry. In 2008, researchers in our unit successfully isolated ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 马冬梅白俊杰邓国成李胜杰叶星于凌云陈昆慈谢俊
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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