Loop-mediated isothermal amplification kit for detecting chicken rhinitis haemophilus paragallinarum and detection method of loop-mediated isothermal amplification kit

A ring-mediated isothermal, Haemophilus gallinus technology, applied in the biological field, can solve problems such as economic loss and difficulty in eradicating completely, and achieve the effects of low detection cost, simple and fast operation, and cost saving

Inactive Publication Date: 2017-11-24
CHONGQING ACAD OF ANIMAL SCI
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  • Abstract
  • Description
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  • 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

Method used

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  • Loop-mediated isothermal amplification kit for detecting chicken rhinitis haemophilus paragallinarum and detection method of loop-mediated isothermal amplification kit
  • Loop-mediated isothermal amplification kit for detecting chicken rhinitis haemophilus paragallinarum and detection method of loop-mediated isothermal amplification kit
  • Loop-mediated isothermal amplification kit for detecting chicken rhinitis haemophilus paragallinarum and detection method of loop-mediated isothermal amplification kit

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[0022] Example 1. Method for detecting pathogenic microorganisms based on loop-mediated isothermal amplification technology

[0023] Based on the method for detecting pathogenic microorganisms based on the loop-mediated isothermal amplification technology, this embodiment takes Haemophilus paragallinarum as an example, and the reagents used are as follows:

[0024] a. The upstream inner primer FIP aqueous solution and the downstream inner primer BIP aqueous solution each with a concentration of 5 μmol / L, the upstream outer primer F3 aqueous solution and the downstream outer primer B3 aqueous solution with a concentration of 20 μmol / L each: the primer sequence is as follows:

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

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

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

[0028] Downstream outer primer B3: 5'-a...

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Abstract

The invention relates to a loop-mediated isothermal amplification kit for detecting chicken rhinitis haemophilus paragallinarum and a detection method of the loop-mediated isothermal amplification kit. The loop-mediated isothermal amplification kit for detecting the chicken rhinitis haemophilus paragallinarum and the detection method of the loop-mediated isothermal amplification kit have the advantages that dextran and astragalus polysaccharides are added into a loop-mediated isothermal amplification system until final concentration is 0.1-0.5mol/L (based on the weight of glucose), so that the stability of polymerase is increased, expensive 'polymerase' and 'betaine' in common LAMP (loop-mediated isothermal amplification) kits can be partially replaced, the amplification of high-content GC fragments can be well promoted, detection reaction time can be reduced to 20-30 minutes, detection cost can be lowered while the detection period can be shortened, and the kit is significant to the loop-mediated isothermal amplification in chicken rhinitis haemophilus paragallinarum detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a loop-mediated isothermal amplification kit for detecting Haemophilus paragallinitis gallinarum, and also relates to a detection method for Haemophilus paragallinitis gallinarum. 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 instruments And reagents, low cost, has shown broad deve...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/21
CPCC12Q1/6844C12Q1/689C12Q2531/119
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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