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Method for quickly detecting pathogenic aeromonas hydrophila by multiple polymerase chain reactions (PCRs)

A technique of Aeromonas hydrophila and pathogenicity, applied in the application field of the method in the diagnosis of Aeromonas hydrophila disease in aquatic animals

Inactive Publication Date: 2012-02-01
中华人民共和国江苏出入境检验检疫局 +1
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AI Technical Summary

Problems solved by technology

A single PCR can only amplify one gene, and the characteristics of a certain pathogen cannot be accurately determined, but multiple PCR can increase its accuracy

Method used

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  • Method for quickly detecting pathogenic aeromonas hydrophila by multiple polymerase chain reactions (PCRs)
  • Method for quickly detecting pathogenic aeromonas hydrophila by multiple polymerase chain reactions (PCRs)
  • Method for quickly detecting pathogenic aeromonas hydrophila by multiple polymerase chain reactions (PCRs)

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Experimental program
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Embodiment 1

[0029] 1. Aseptically take the bacteria with Aeromonas hydrophila ( A. hydrophila) (Refer to Chopra A.K., Houston C.W., Peterson J.W. & Jin G.-F. (1993) Cloning, expression, and sequence analysis of a cytolytic enterotoxin gene from Aeromonas hydrophila. Canadian Journal of Microbiology 39, 513–523 for specific operations and identification. , identification of strains artificially) Infected with typical clinical symptoms of heterogeneous gibel crucian carp ( Carassius auratus gibelio ) 10g of festered muscle tissue, homogenized with a homogenizer. Take 1g of the homogenate and put it into a 1.5ml centrifuge tube, centrifuge at 10000g for 10min, discard the supernatant, resuspend the precipitate with distilled water, centrifuge at 10000g for 10min, add 1ml of distilled water to resuspend, cover the centrifuge tube, boil in boiling water for 10min, centrifuge at 10000g After 10 minutes, the supernatant was transferred to a new sterile centrifuge tube as a DNA template for mu...

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Abstract

The invention relates to a method for quickly detecting pathogenic aeromonas hydrophila by multiple polymerase chain reactions (PCRs). The method comprises the steps of: designing three pairs of primers by using aeromonas hydrophila 16rDNA (Ribosomal deoxyribonucleic acid) gene conservation region sequence, ectotoxin gene and quorum-sensing signal gene sequence, and building the quick detection method by the multiple PCRs. The method is high in specificity, convenient to operate, low in detection cost and short in time, can be used for detecting a large batch of samples at the same time and can be standardized into a product, thus meeting the demand of an inspection and quarantine department which needs to detect the pathogenic aeromonas hydrophila of aquatic animals on a large scale.

Description

[0001] technical field [0002] The invention belongs to the field of biotechnology, and relates to a molecular biology method for rapidly detecting pathogenic Aeromonas hydrophila, and also relates to the application of the method in the diagnosis of Aeromonas hydrophila disease in aquatic animals. Background technique [0003] With the continuous expansion of the scale of intensive farming of aquatic animals, the occurrence of diseases is becoming more and more frequent, which seriously restricts the healthy development of aquaculture industry. There are many pathogens of aquatic animal diseases, and the traditional isolation and culture of microorganisms for identification can no longer meet the needs of aquatic animal disease prevention and control. Therefore, the establishment of rapid, accurate, and sensitive pathogen detection methods is the prerequisite for timely control and prevention of diseases. Traditional microbiological detection methods are time-consuming a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
Inventor 姜焱储卫华王凯民陈雷朱卫侯玉峰张常印
Owner 中华人民共和国江苏出入境检验检疫局
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