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Kit for rapidly detecting Edwardsiella ictalurus of fishes and detection method

A detection method and kit technology, applied in the biological field, can solve the problems of long time, high cost, and high equipment requirements, and achieve the effects of low cost, improved economic benefits and food safety.

Pending Publication Date: 2020-06-12
NEIJIANG NORMAL UNIV
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  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the above-mentioned problems existing in the prior art, the present invention provides a kit and a detection method for the rapid detection of Edwardsiella spp., which can effectively solve the problems of high cost, complicated operation and high equipment requirements of the existing diagnostic methods. and time-consuming problems

Method used

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  • Kit for rapidly detecting Edwardsiella ictalurus of fishes and detection method
  • Kit for rapidly detecting Edwardsiella ictalurus of fishes and detection method
  • Kit for rapidly detecting Edwardsiella ictalurus of fishes and detection method

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Embodiment 1

[0030] A kit for rapid detection of Edwardsiella catfish, comprising: polylysine-treated slides (solid-phase carrier slides), rabbit polyclonal antibody against Edwardsiella catfish, HRP marker goat anti-rabbit IgG, blocking solution and chromogenic solution.

[0031] Wherein, the blocking solution is BSA with a mass concentration of 5%, and the color developing solution is TMB solution.

[0032] Rabbit polyclonal antibody against Edwardsiella spp. was prepared by the following method:

[0033] (1) Inoculate Edwardsiella spp. on a BHI plate, culture it in an incubator at 27°C for 24 hours, check its purity by Gram staining method, and sequence it by PCR. After confirming that it is Edwardsiella and no other bacteria, use 0.6 % formaldehyde to inactivate bacteria, prepared at a concentration of 1 x 10 8 cfu / ml inactivated vaccine;

[0034] (2) Use the inactivated vaccine prepared in step (1) to immunize New Zealand white rabbits, immunize 4 times, and immunize once every oth...

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Abstract

The invention provides a kit for rapidly detecting Edwardsiella ictalurus of fishes and a detection method. The kit comprises a slide treated by polylysine, a rabbit anti-Edwardsiella ictalurus polyclonal antibody, an HRP labeled goat anti-rabbit lgG, a confining liquid and a developing liquid. The slide is used as a solid-phase carrier, so that the detection cost is greatly reduced; the rabbit anti-Edwardsiella ictalurus polyclonal antibody prepared by the invention can recognize a plurality of epitopes of the same antigen, and is strong in specificity and high in sensitivity; TMB is used asa peroxidase substrate, TMB is used as a developing solution, a soluble light blue product can be generated, and the light blue product is adsorbed on the filter paper, so that the color change can bevisually observed by naked eyes. The detection method is low in equipment requirement, low in cost, simple and convenient to operate, green, environment-friendly and high in sensitivity.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and a detection method for rapidly detecting the fish Edwardsiella chrysalis. Background technique [0002] With the development of the economy, people's pursuit of nutrients is more intense. Fish is rich in protein and is a good source of nutrition, especially pelteobagrus fulvidraco. Pelteobagrus fulvidraco belongs to the order Siluriformes, the family Pelteidae, and the genus Pelteobagrus. It is a high-quality and precious freshwater fish in my country, commonly known as yellow drum fish , Huang Lading and so on. Yellow catfish is rich in protein, which has the effects of maintaining potassium and sodium balance, eliminating edema, improving immunity, lowering blood pressure, and buffering anemia, which is beneficial to the growth and development of teenagers. At the same time, yellow catfish is rich in copper. Copper is an indispensable trace nutrient for human...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/535
CPCG01N33/56911G01N33/535G01N2333/195Y02A40/81
Inventor 贺扬王均陈杰姜静王成鑫向捷刘用平黎金玉
Owner NEIJIANG NORMAL UNIV
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