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Application of dextran-combined astragalus polysaccharide in detection of haemophilus paragallinarum causing infectious rhinitis of chicken as well as kit and detection method of dextran-combined astragalus polysaccharide

A technology of Haemophilus gallinarum and dextran, which is applied in the field of kits containing dextran and astragalus polysaccharides, can solve the problems of difficult and complete eradication, economic loss, etc., and achieve simple and fast operation, low detection cost and strong specificity Effect

Inactive Publication Date: 2017-11-24
CHONGQING ACAD OF ANIMAL SCI
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  • Description
  • Claims
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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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  • Application of dextran-combined astragalus polysaccharide in detection of haemophilus paragallinarum causing infectious rhinitis of chicken as well as kit and detection method of dextran-combined astragalus polysaccharide
  • Application of dextran-combined astragalus polysaccharide in detection of haemophilus paragallinarum causing infectious rhinitis of chicken as well as kit and detection method of dextran-combined astragalus polysaccharide
  • Application of dextran-combined astragalus polysaccharide in detection of haemophilus paragallinarum causing infectious rhinitis of chicken as well as kit and detection method of dextran-combined astragalus polysaccharide

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Embodiment 1

[0023] Embodiment 1, method for detecting pathogenic microorganisms based on loop-mediated isothermal amplification technology

[0024] 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:

[0025] 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:

[0026] Upstream internal primer FIP: 5'-acttgacgtcatccccaccttcgggtgggaactcaaaggag-3' (SEQ ID NO.1);

[0027] Downstream internal primer BIP: 5'-cgagtagggctacacacgtgctgcactttctgagattcgct-3' (SEQ ID NO.2);

[0028] Upstream outer primer F3: 5'-tcctttgttgccagcacttc-3' (SEQ ID NO.3);

[0029]Downstream oute...

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Abstract

The invention relates to application of dextran-combined astragalus polysaccharide in detection of haemophilus paragallinarum causing infectious rhinitis of chicken as well as a kit and a detection method of the dextran-combined astragalus polysaccharide. By adding dextran and astragalus polysaccharide in a loop-mediated isothermal amplification system until the final concentration is 0.1-0.5mol / L (calculated by glucose), the stability of polymerase is improved, expensive 'polymerase' and 'glycine betaine' in a common LAMP kit can be partly replaced, a high-content GC fragment can be promoted to amplify very well, the detection reaction time can be reduced to 20-30 minutes, the detection cost can be reduced, meanwhile, the detection period is shortened, and the dextran-combined astragalus polysaccharide has important significance for loop-mediated isothermal amplification in the detection of the haemophilus paragallinarum causing the infectious rhinitis of chicken.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of dextran combined with astragalus polysaccharide in loop-mediated isothermal amplification detection of Haemophilus paragallinitis gallinarum, and also relates to a kit containing dextran and astragalus polysaccharide and a detection method. 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 ope...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/21
CPCC12Q1/6844C12Q2531/119C12Q2527/125C12Q2563/107
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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