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Gene chip capable of simultaneously detecting various vibrios and method for detecting vibrios

A gene chip and gene technology, applied in the direction of microbe-based methods, biochemical equipment and methods, microbiological measurement/testing, etc., to achieve efficient and fast detection, high specificity, and good stability

Inactive Publication Date: 2013-12-18
THE FIRST INST OF OCEANOGRAPHY SOA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The specificity and sensitivity of immune antibody detection technology are high, but often only a single bacterial species can be tested at a time, and antiserum needs to be prepared

Method used

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  • Gene chip capable of simultaneously detecting various vibrios and method for detecting vibrios
  • Gene chip capable of simultaneously detecting various vibrios and method for detecting vibrios
  • Gene chip capable of simultaneously detecting various vibrios and method for detecting vibrios

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Embodiment Construction

[0041] 1. Probe Design

[0042] The gene chip of the invention is used to detect vibrio alginolyticus, vibrio parahaemolyticus, vibrio harveyi and vibrio vulnificus in fishery breeding environment and aquatic products. The probes used are all designed according to the hypervariable region sequences of the hsp60 and toxR genes of the strain, and their specificity is guaranteed by BLAST comparison. table 3. The primers and probes used were commercially synthesized by specialized companies.

[0043] Table 3. Sequence comparison information of the probes applied for in the present invention and similar probes of other published patents

[0044]

[0045] Note: Approximate probe patent publication numbers: 1. CN101475986A; 2. CN101475986A; 3. CN101967510A; 4. CN101967510A; 5. CN101113476A.

[0046] 2. The construction of the gene chip:

[0047] (1) Dilute the commercially synthesized oligonucleotide probe to an appropriate concentration: use 1×TE buffer to dilute to a concent...

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Abstract

The invention relates to a gene chip capable of simultaneously detecting various vibrios and a method for detecting the vibrios, and provides species-specific genes of four common pathogenic vibrios in the aquatic environment to encode an oligonucleotide probe sequence of an hsp60 gene and an oligonucleotide probe sequence of a toxR gene, wherein the four common pathogenic vibrios include vibrio alginolyticus, vibrio parahaemolyticus, vibrio vulnificus and vibrio haeveyi. On the basis of the base complementary and pairing principle, by combining the method with an optimized ordinary PCR and fluorescence labeled PCR reaction system, the aim of detection is achieved through oligonucleotide probes fixed to a solid-phase aldehyde chip. The invention provides the detecting method which is high in flux, specificity and sensitivity, rapid and efficient for the bottleneck of a current method for detecting pathogenic microorganisms.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a gene chip capable of parallel detection of various Vibrio and a detection method thereof. Background technique [0002] There are many methods for microbial detection. At present, traditional physiological and biochemical experimental methods, immune antibody detection technology, PCR detection technology, etc. are widely used. Traditional physiological and biochemical experiments need to isolate and cultivate the strains of the samples to be tested, and identify the strains through a series of tedious characteristics and results of physiological and biochemical experiments. Although this method is a commonly used and classic identification method in microbiology laboratories, However, the process often takes several days, requires a lot of manpower and material resources, and the phenotypic characteristics obtained by the experiment may be affected...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C40B40/06C40B50/18C12R1/63
Inventor 曲凌云朱鹏飞李壹田欣欣王琛
Owner THE FIRST INST OF OCEANOGRAPHY SOA
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