Reagent for detection of pathogenic hydrophila gingivalis and detection method thereof

A technology for detection of Aeromonas hydrophila and a detection method, which is applied in the field of reagents for detection of pathogenic Aeromonas hydrophila, can solve problems such as time-consuming, cumbersome operation, and difficulty in grasping, and achieve simple operation, strong specificity, easy to get effect

Inactive Publication Date: 2009-04-29
INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method is relatively cumbersome to operate, and the data processing of the experimental results requires artificial comparison and judgment

Method used

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  • Reagent for detection of pathogenic hydrophila gingivalis and detection method thereof
  • Reagent for detection of pathogenic hydrophila gingivalis and detection method thereof
  • Reagent for detection of pathogenic hydrophila gingivalis and detection method thereof

Examples

Experimental program
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Effect test

Embodiment Construction

[0021] Randomly take 6 water samples from the water body of a breeding pond of a certain eel factory in Fujian Province, separate the pathogenic bacteria, take 30ml water samples each, centrifuge, 1000rpm (1000 revolutions per minute) for 5mins, take the supernatant, 3900rpm Centrifuge for 30mins, discard the supernatant, suspend the precipitate with 2ml TE buffer, take a small amount of the suspension in a test tube containing 5ml sterilized alkaline peptone water, incubate at 30°C for 15 hours, then streak and inoculate on an ordinary agar plate, incubate at 30°C for 15 hours Hour.

[0022] Then carry out strain culture: the standard strain of Aeromonas hydrophila was cultured in common broth based on 30° C., 200 rpm shaking culture for 15 hours.

[0023] Preparation of template DNA: Take 1.5ml of the above-mentioned bacterial solution, separate it with a centrifugal force of 15000g for 10 minutes, and collect the bacterial precipitate. Suspend the bacterial pellet with 150...

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Abstract

The invention discloses a reagent for detecting pathogenic aeromonas hydrophila, which comprises the following solutions: 1 milliliter of 10xPCR buffer solution, 60 microliters of Pf solution, 60 microliters of Pr solution, 1 milliliter of 2xPCR water and 30 microliters of Taq enzyme. The invention also relates to a detection method which applies the reagent for detecting the pathogenic aeromonas hydrophila. The reagent has the characteristics of high sensitivity, strong specificity and easily obtained template materials, and the detection method has the characteristics of simple and convenient operation and quickness and time conservation.

Description

technical field [0001] The invention relates to a reagent for detecting pathogenic Aeromonas hydrophila and a detection method thereof. Background technique [0002] Aeromonas hydrophila (A.hydrophila) widely exists in fresh water and water containing organic matter, silt and fresh water fish body surface. Aeromonas hydrophila can be divided into pathogenic strains and non-pathogenic strains. Pathogenic Aeromonas hydrophila can produce highly toxic exotoxins, such as: hemolysin (hemolysin), tissue toxin, necrotic toxin, enterotoxin and protease. Pathogenic strains can infect animals such as fish, amphibians, reptiles, birds and mammals. It can cause a variety of fish infectious diseases, especially red fin disease, fulminant hemorrhagic disease, and sepsis, which cause a large number of eels, and the mortality rate is very high. At the same time, Aeromonas hydrophila can also cause human diseases, such as acute gastroenteritis in people of different ages. The current met...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 李寿崧林俊生江树勋饶静静江云陈文炳邵碧英黄晓蓉
Owner INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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