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Gene chip and applications thereof in detection of aquatic pathogenic microorganism

A gene chip and gene technology, applied in the field of aquatic pathogenic microorganism detection gene chip, can solve the problem of time-consuming and labor-intensive

Inactive Publication Date: 2011-08-17
HANGZHOU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] An object of the present invention is to provide a rapid detection of aquatic pathogenic microorganisms such as Aeromonas hydrophila, Edwardsiella tarda, Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella, Shigella, etc. Gene chips to overcome the time-consuming and labor-intensive defects of existing technologies, improve detection sensitivity and specificity, and shorten the detection cycle

Method used

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  • Gene chip and applications thereof in detection of aquatic pathogenic microorganism
  • Gene chip and applications thereof in detection of aquatic pathogenic microorganism
  • Gene chip and applications thereof in detection of aquatic pathogenic microorganism

Examples

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Embodiment 1

[0028] The establishment of embodiment 1 Aeromonas hydrophila PCR detection system

[0029] Aeromonas hydrophila (Aeromonas hydrophila) belongs to Aeromonas family (Aermonadaceae) Aeromonas (Aeromonas), is an opportunistic pathogenic bacteria shared by humans, livestock and aquatic animals. The bacterium widely exists in the water environment and is the main pathogenic bacterium of various aquatic animals. Aeromonas hydrophila is an opportunistic pathogen that widely exists in nature, especially in water environments. The virulence of Aeromonas hydrophila varies greatly, and non-toxic strains exist widely. Traditional bacterial isolation and identification must be combined with biological or serological methods of toxins to determine its pathogenicity, which is time-consuming, laborious and not sensitive; due to the existence of aer + hp - genotype and aer - hp + For pathogenic strains of genotype, the PCR method established for aer or a such as a single gene may have mis...

Embodiment 2

[0044] Example 2 Establishment of Vibrio parahaemolyticus PGR detection system

[0045] Vibrio parahaemolyticus is one of the important pathogens causing food poisoning in aquatic products, and it is an important indicator in food safety testing. In my country's coastal areas, food poisoning caused by Vibrio parahaemolyticus accounts for the first place in bacterial food poisoning incidents. The statutory detection method of Vibrio parahaemolyticus is still the traditional culture method, which is cumbersome, time-consuming and laborious, and often takes 5-6 days to complete.

[0046] The main virulence factors of Vibrio parahaemolyticus include a variety of hemolytic toxins, mainly thermolabile direct hemolytic toxin (TLH), thermotolerant direct hemolytic toxin (TDH) and thermotolerant direct hemolytic toxin (TRH), which are composed of tlh, tdh and trh genes encode. The Tlh ​​gene exists in all Vibrio parahaemolyticus and is species-specific. Therefore, specific primers t...

Embodiment 3

[0047] The establishment of other aquatic pathogenic bacteria PCR detection system of embodiment 3

[0048] With reference to the ideas in Examples 1 and 2, by optimizing the PCR amplification conditions and perfecting the design of primers, the PCR detection system for common aquatic pathogenic microorganisms such as Edwardsiella tarda was determined. The amplification conditions are shown in Table 6.

[0049] Table 6 PCR amplification primers for common aquatic pathogenic microorganisms

[0050]

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Abstract

The invention relates to a gene chip in the detection of aquatic pathogenic microorganism, a preparation method and applications thereof in the detection of aquatic pathogenic microorganism. A conserved sequence selecting from a target pathogenic microorganism gene is fixed on a solid-phase carrier as an oligonucleotide probe, in the proper hybridization condition, a marked sample to be detected is added for reaction, and the corresponding aquatic pathogenic microorganism can be identified according to a probe display signal on the gene chip. The gene chip can fast detect the aquatic pathogenic microorganism such as aeromonas hydrophila, edwardsiella tarda, staphylococcus aureus, vibrio parahaemolyticus, listeria monocytogenes, salmonella typhimurium, shigella and the like, thus being high in detection sensitivity, and strong in specificity and greatly shortening the detection period.

Description

technical field [0001] The invention relates to a gene chip for detecting aquatic pathogenic microorganisms, its preparation method and its application in detecting aquatic pathogenic microorganisms. Background technique [0002] Since the reform and opening up, my country's fishery has achieved world-renowned achievements. The total output of aquatic products has been ranking first in the world since 1990, and it has developed into a world fishery power. Fishery plays an increasingly important role in my country's national economy, especially in the development of agricultural economy. status. [0003] However, the destruction of the ecological environment, the decline in growth rate, and the outbreak of a large number of diseases have become problems that cannot be ignored in the aquaculture industry and need to be solved urgently. In recent years, grass carp haemorrhagic disease, crustacean white spot syndrome and Taura syndrome, freshwater fish bacterial sepsis and soft-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCY02A50/30
Inventor 许宝青冯晓宇郭水荣林启存蔡丽娟刘凯
Owner HANGZHOU ACAD OF AGRI SCI
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