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Kit for detecting chicken coryza haemophilus paragallinarum on basis of loop-mediated isothermal amplification and detection method of kit

A ring-mediated isothermal, Haemophilus gallinarum technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as economic loss, difficulty in eradicating it completely, and achieve low detection cost, simple and rapid operation, Guaranteed effect of reliability

Inactive Publication Date: 2017-11-07
CHONGQING ACAD OF ANIMAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Once Haemophilus paragallinitis infection occurs in chicken flocks in a certain area, the disease will become an endemic disease, which is difficult to eradicate completely, and often recurs, causing huge economic losses

Method used

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  • Kit for detecting chicken coryza haemophilus paragallinarum on basis of loop-mediated isothermal amplification and detection method of kit
  • Kit for detecting chicken coryza haemophilus paragallinarum on basis of loop-mediated isothermal amplification and detection method of kit
  • Kit for detecting chicken coryza haemophilus paragallinarum on basis of loop-mediated isothermal amplification and detection method of kit

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Embodiment 1

[0022] Embodiment 1, method for detecting pathogenic microorganisms based on loop-mediated isothermal amplification technology

[0023] Based on the method of loop-mediated isothermal amplification technology to detect pathogenic microorganisms, the present embodiment takes Haemophilus paragallinitis as an example, and the reagents used are as follows:

[0024] a. The upstream internal primer FIP aqueous solution and the downstream internal primer BIP aqueous solution with a concentration of 5 μmol / L, the upstream external primer F3 aqueous solution and the downstream external primer B3 aqueous solution with a concentration of 20 μmol / L: the primer sequences are as follows:

[0025] Upstream internal primer FIP: 5'-gcacgtgtgtagccctactcgctggaggaaggtggggatg-3' (SEQ ID NO.1);

[0026] Downstream internal primer BIP: 5'-ggtgcatacagagggaagcgaggttgcagactccaatccgg-3' (SEQ ID NO.2);

[0027] Upstream outer primer F3: 5'-caaaggagactgccagtgat-3' (SEQ ID NO.3);

[0028] Downstream oute...

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Abstract

The invention relates to a kit for detecting chicken coryza haemophilus paragallinarum on the basis of loop-mediated isothermal amplification and a detection method of the kit. By adding dextran and astragalus polysaccharide in a loop-mediated isothermal amplification system to reach the final concentration being 0.1-0.5 mol / L (on the basis of glucose), the stability of polymerase is improved, polymerase and betaine which are high in price in an ordinary LAMP kit can be partially replaced, amplification of high content GC fragments can be well promoted, the detection reaction time can be shortened to 20-30 minutes, and the detection period can be shortened while the detection cost is lowered; an important meaning to loop-mediated isothermal amplification on chicken coryza haemophilus paragallinarum detection is achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for detecting Haemophilus paragallinitis gallinarum based on loop-mediated isothermal amplification, and also relates to a method for detecting Haemophilus paragallinitis gallinarum based on loop-mediated isothermal amplification. Background technique [0002] At present, the detection methods of original microorganisms mainly include isolation and culture identification, polymerase chain reaction (PCR) and fluorescence quantitative PCR (FQ-PCR). The method requires expensive instruments and reagents, and the detection cost is high, so it is not suitable for small and medium-sized units and on-site detection applications. [0003] Compared with other nucleic acid amplification techniques, loop-mediated isothermal amplification (LAMP) technology can rapidly, efficiently and specifically amplify target sequences under isothermal conditions, and is easy to operate without expensive ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6844C12Q1/6848C12Q1/689C12Q2531/119C12Q2527/125C12Q2563/107
Inventor 杨金龙殷素会戴茜茜许国洋周婵王海燕谷山林王介平吕金凤王小燕李晋
Owner CHONGQING ACAD OF ANIMAL SCI
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