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Aeromonas hydrophila aerolysin Dot-ELISA detection method

A technology for the detection of Aeromonas hydrophila and its detection method, applied in the field of Dot-ELISA detection of pathogenic Aeromonas hydrophila, which can solve the problems of difficult standardization and complex components of the extracellular products of Aeromonas hydrophila, etc. Achieve intuitive results, fast and effective detection, and good reliability

Inactive Publication Date: 2013-04-10
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
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Problems solved by technology

[0004] The Dot-ELISA established for the extracellular products of Aeromonas hydrophila has been included in the national standard for detection methods of pathogenic Aeromonas hydrophila (GB / T 186522-2002), but because the extracellular products of Aeromonas hydrophila The composition is relatively complex, including extracellular toxins such as aerolysin, hemolysin and cytotoxic enterotoxin, as well as extracellular proteases, ribozymes and amylases, so the Dot-ELISA detection method based on total extracellular products is not easy to standardize

Method used

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  • Aeromonas hydrophila aerolysin Dot-ELISA detection method

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Embodiment Construction

[0032] The following is in conjunction with specific embodiments. The experimental method of unreceipted specific conditions in the embodiment, usually adopts conventional conditions, such as the condition described in " molecular cloning laboratory manual " (New York: Cold Spring Harbor Laboratory Press, 1989) (people such as Sambrook), or according to The method recommended by the manufacturer.

[0033] (1) Establishment of a Dot-ELISA method for the detection of Aeromonas hydrophila aerolysin

[0034] 1. Selection of standard strains

[0035] The standard strain used in the present invention is the J-1 strain of Aeromonas hydrophila, which is classified and named: Aeromonas hydrophila. The depository management committee general microbiology center (CGMCC), its deposit number is CGMCC NO.3220.

[0036] 2. Prokaryotic expression and purification of recombinant aerolysin from Aeromonas hydrophila

[0037] 1) Cloning of Aeromonas hydrophila aerolysin gene

[0038] The tot...

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Abstract

The invention provides an aeromonas hydrophila recombinant aerolysin Dot-enzyme-linked immuno sorbent assay (ELISA) detection method and belongs to the technical field of biology. A nitrocellulose (NC) film is taken as a solid-phase carrier, aeromonas hydrophila recombinant aerolysin rabbit antiserum is taken as a primary antibody, and a hypothalamic regulatory peptide (HRP) goat-anti-rabbit IgG is taken as a secondary antibody. The method can measure a plurality of samples, can obviously shorten the time for detecting the samples, can be operated easily and rapidly, can meet the requirement under a non-laboratory condition simultaneously, and can be rapidly and conveniently used for detecting pathogenic aeromonas hydrophila.

Description

technical field [0001] The invention relates to a method in the field of biotechnology, in particular, to a method for Dot-ELISA detection of pathogenic Aeromonas hydrophila. Background technique [0002] Aeromonas hydrophila belongs to the genus Aeromonas of the Vibrio family, and is a mesophilic, motile group of Aeromonas that is ubiquitous in fresh water, sewage, silt, soil and human feces . Pathogenic Aeromonas hydrophila is pathogenic to aquatic animals, livestock and humans, and can not only cause gill rot, red skin disease, perforation and other diseases of aquatic animals, but also can pass through aquatic animals and aquatic products. It infects humans and animals, causing diarrhea and food and feed poisoning in humans and animals. It is a typical human-animal-fish pathogen. In view of the extensive pathogenicity of Aeromonas hydrophila, its rapid and effective diagnosis, epidemiological investigation and public health quarantine are of great value. [0003] The ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569C12N15/31C12N15/70C07K14/28C07K16/12C07K16/06
Inventor 刘永杰陆承平焦大为
Owner NANJING AGRICULTURAL UNIVERSITY
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