Application of astragalus polysaccharide in loop-mediated isothermal amplification, loop-mediated isothermal amplification detection kit containing astragalus polysaccharides and detection method

A ring-mediated isothermal and astragalus polysaccharide technology, applied in the field of kits containing astragalus polysaccharides, can solve problems such as economic loss, foul smell, and difficulty in eradicating them completely, and achieve low detection cost, short time-consuming, and shortened detection cycle. Effect

Pending Publication Date: 2017-11-24
CHONGQING ACAD OF ANIMAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Once Klebsiella gallinarum infection occurs in chicken flocks in a certain area, the disease will become an endemic disease, it is difficult to completely eradicate, and frequent repeated outbreaks will bring huge economic losses
The main pathological features of Klebsiella chicken rhinitidis infection are chronic atrophic rhinitis, stench, sepsis, and urinary tract infection. It is difficult to distinguish it from Haemophilus paragallinarum infection pathologically.

Method used

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  • Application of astragalus polysaccharide in loop-mediated isothermal amplification, loop-mediated isothermal amplification detection kit containing astragalus polysaccharides and detection method
  • Application of astragalus polysaccharide in loop-mediated isothermal amplification, loop-mediated isothermal amplification detection kit containing astragalus polysaccharides and detection method
  • Application of astragalus polysaccharide in loop-mediated isothermal amplification, loop-mediated isothermal amplification detection kit containing astragalus polysaccharides and detection method

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Embodiment 1

[0026] Embodiment 1, method for detecting pathogenic microorganisms based on loop-mediated isothermal amplification technology

[0027] Based on the method of loop-mediated isothermal amplification technology to detect pathogenic microorganisms, the present embodiment takes Klebsiella gallinarum as an example, and the reagents used are as follows:

[0028] 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:

[0029] Upstream inner primer FIP: 5'-gcacatctgatggcatgaggcc-aataccgcataacgtcgca-3' (SEQ ID NO.1);

[0030] Downstream internal primer BIP: 5'-tggggtaatggctcacctaggc-agtgtggctggtcatcctc-3' (SEQ ID NO.2);

[0031] Upstream outer primer F3: 5'-ggagggggataactactgga-3' (SEQ ID NO.3);

[0032] Downstream outer prime...

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Abstract

The invention relates to application of astragalus polysaccharides in loop-mediated isothermal amplification, a loop-mediated isothermal amplification detection kit containing the astragalus polysaccharides and a detection method. The application of the astragalus polysaccharides in the loop-mediated isothermal amplification, the loop-mediated isothermal amplification detection kit containing the astragalus polysaccharides and the detection method have the advantages that the 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 application is significant to the loop-mediated isothermal amplification in pathogenic microorganism detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the application of astragalus polysaccharide in ring-mediated isothermal amplification, and also relates to a kit containing 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 operate without expensive instruments And reagents, low cost, has shown broad development p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12R1/22
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2527/125C12Q2563/107
Inventor 杨金龙戴茜茜殷素会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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