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PCR test kit for hygrophilous aeromonad and its test method

A technology of Aeromonas hydrophila and detection kit, which is applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of complex biochemical detection operation process, high technology and equipment conditions, and long detection cycle. , to achieve the effect of eliminating false positive reactions, simple experimental equipment, and shortened detection cycle

Inactive Publication Date: 2004-06-23
福建省农业科学院生物技术中心
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Problems solved by technology

[0004] At present, the detection of Aeromonas hydrophila is mainly based on conventional biological detection methods, which have the following disadvantages: there are many false positives and false negatives in the detection process; the biochemical detection operation process is complicated, and the detection cycle is long. It takes two to three days to test the sample to be tested once; biochemical testing usually requires the strain to be tested to be cultured for a long time before testing; biochemical testing requires high technology and equipment conditions, and the cost of testing is high

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  • PCR test kit for hygrophilous aeromonad and its test method
  • PCR test kit for hygrophilous aeromonad and its test method
  • PCR test kit for hygrophilous aeromonad and its test method

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Embodiment Construction

[0065] The present invention will be described in more detail below in conjunction with examples.

[0066] Best practice:

[0067] 1. Preparation of materials and equipment required for testing experiments

[0068] 1. Kit composition:

[0069] 1)MgCl 2 (25mM) 50μl

[0070] 2) dNTP (2mM) 50μl

[0071] 3) 10×buffer (10×enzyme-specific reaction buffer)

[0072] 100μl

[0073] 4) Taq polymerase (3U / μl) (Taq polymerase) 10μl

[0074] 5) Primer mixture (primer) (50pmol) 30μl

[0075] 6) Aeromonas hydrophila positive control (CK + ) 20μl

[0076] 7) Aeromonas hydrophila negative control (CK - ) 20μl

[0077] 8)ddH 2 O 1ml

[0078] Each kit can be used to detect 20 samples.

[0079] where MgCl 2 , 10×buffer, and Taq polymerase were provided by Huamei Bioengineering Company; dNTP was provided by Shanghai Sangon Bioengineering Technology Service Co., Ltd.; Primermixture was provided by...

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Abstract

The PCR test kit for detecting hygrophilous aeromonad includes primer, dNTP, buffering solution and DNA polymerase, and its upstream primer is 5-CCAAGGGGTCTGTGGCGACA-3í» and downstream primer is 5í»-TTTCACCGGTAACAGGATTG-3í». The PCR test method includes providing sample template; mixing in PCR thin-wall tube dNTP, MgCl2, buffer, primer, DNA, polymerase, tested sample and ddH2O; amplifying the mixture in PCR instrument; electrophoresing the amplified product in electrophoresis equipment while recording result; and analysis and judgment. The PCR test kit is easy to produce and the test method has high sensitivity, short test period, high test precision reaching 10 aeromonad or 1 ng template DNA, and relatively low test cost.

Description

technical field [0001] The invention relates to a composition used in the measurement or inspection of microorganisms and a detection method thereof, in particular to a PCR kit for detecting Aeromonas hydrophila and how to use the kit to detect Aeromonas hydrophila. Background technique [0002] Aeromonas hydrophila (Aeromonas hydrophila) belongs to the Vibriaceae Aeromonas genus, is a pathogenic bacteria of fish, amphibians, reptiles and mammals, and is also a high-value aquatic animal such as Chinese soft-shelled turtle and eel The main pathogen of fulminant infectious diseases, the toxin produced by the bacteria can cause comprehensive symptoms such as animal rotting skin, perforation, ulcer, bleeding, sepsis, etc., thus indirectly causing human diarrhea. [0003] In order to reduce or prevent this disease, it is generally necessary to detect Aeromonas hydrophila on susceptible animals so that animals infected with the pathogen can be isolated and treated, or the cause ca...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/25C12Q1/68
Inventor 宋铁英郑伟文
Owner 福建省农业科学院生物技术中心
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