Aquatic pathogenic bacterium colony induction signaling molecule detecting method and special reagent kit thereof

A technology of quorum sensing signals and detection kits, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, analysis through chemical reactions of materials, etc., and can solve the limitations of single biological reporting system, detection range and accuracy , false negative and other problems, to achieve the effect of rapid results, low cost and high accuracy

Inactive Publication Date: 2007-11-07
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

However, the published biological reporting system is relatively simple (Zhan Kehang, et al. Chinese Medical Journal. 86(37): 2655-2658, 2006), there are certain limitations in the detection range and accuracy, and

Method used

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Examples

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Embodiment 1

[0040] Example 1. A method for rapid detection of quorum sensing signal molecules of aquatic gram-negative pathogens

[0041] 1. Preparation of a rapid detection kit for quorum sensing signal molecules of aquatic gram-negative pathogens

[0042] 1) Cultivation of report strains

[0043] The above reporter strain Agrobacterium tumefaciens A136 was added to 4.5μg / ml (final concentration) tetracycline, 50.0μg / ml (final concentration) spectinomycin LB solid medium (1% peptone, 0.5% yeast extract (0.5% NaCl, 1.2% agar), cultured at 25°C for 1 day;

[0044] Chromobac terler violaceum CV026 was streaked on LB solid medium supplemented with 20.0μg / ml (final concentration) kanamycin, and cultured at 25°C for 1 day;

[0045] E. coli (Escherichia coli) pSB403 added tetracycline 20.0μg / ml (final concentration) LB solid medium (1% peptone, 0.5% yeast extract, 0.5% NaCl, 1.2% agar) was streaked and cultured at 25°C 1 day.

[0046] 2) Cultivation of positive control strains

[0047] Positive con...

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Abstract

The invention discloses an examination method of aquatic product disease germ community induce signal member and its special-purpose reagent box. The method including carrying on the following experiment of the received test fungus: 1) adds X-Gal or ONPG on the plate with the root cancer farming bacillus together streak cultivation, observes whether demonstrates the blue color; 2) with the purple bacillus CV026 on the identical plate streak cultivation, observes whether demonstrates the purple color; 3) puts the received test fungus score inoculate on the LB plate which contains 500nM N-acylation homoserine lactone cultivated, observes whether demonstrated the purple discoloration will be shallow or removes; 4) draws a line together with Esc.coli pSB403 on the plate cultivated, observes whether shines; in above experiment, if at least an experiment demonstrates the masculine gender, then the received test fungus produces the N- acylation homoserine lactone. The invention method has high accurate rate, have no false negative phenomenon, easy to qualitative and quantitative analysis, operation simple, cost low, rapid to obtain the result, suits the large-scale high efficiency examination.

Description

Technical field [0001] The invention relates to a rapid detection method for quorum sensing signal molecules of aquatic gram-negative pathogenic bacteria and a special kit thereof. Background technique [0002] The economic loss of aquaculture caused by bacterial diseases in China is as high as 50 billion yuan each year. The pathogenic bacteria of aquatic products are mainly Gram-negative bacteria (Jiang Lan, et al. China Fisheries Science. 9(2): 152-156, 2002), common These include Vibrio anguillarum, Aeromonas salmonicida, Aeromonas hydrophila, Yersiniaruckeri, Vibrio salmonicida, Vibrio vulnificus, Aeromonas salmonicida and Vibrio splendidus (Jesper BBet al. Dis Aquat Org. 65: 43-52, 2005), which are all related to water environmental pathogens. It is related and should be distinguished from the more researched plant pathogens (Gao Yijing, Nanjing Agricultural University master's degree thesis, the effect of the quorum sensing system of the mesophytic rhizobia on its growth an...

Claims

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Application Information

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IPC IPC(8): G01N21/00G01N21/25G01N21/76G01N30/90C12Q1/04
Inventor 周志刚姚斌何夙旭刘玉春石鹏君罗会颖杨培龙孟昆
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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