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Method for detecting anaplasma phagocytophilum employing loop-mediated isothermal amplification (LAMP) technology

A phagocytic and technical detection technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of low detection rate and cumbersome operation of detection methods, and achieve simple result judgment, simple operation and sensitivity. high effect

Inactive Publication Date: 2014-04-02
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0008] Aiming at the problems of low detection rate and cumbersome operation of the detection methods in the prior art, the present invention provides a method for detecting phagophilia anplasma using LAMP technology. Rapid, sensitive, efficient, low-cost and other characteristics provide a scientific basis for the rapid diagnosis of aplasmosis

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  • Method for detecting anaplasma phagocytophilum employing loop-mediated isothermal amplification (LAMP) technology
  • Method for detecting anaplasma phagocytophilum employing loop-mediated isothermal amplification (LAMP) technology
  • Method for detecting anaplasma phagocytophilum employing loop-mediated isothermal amplification (LAMP) technology

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Embodiment Construction

[0032] The present invention is described in further detail below by embodiment.

[0033] The reagents are all commercially available, and the DNA templates used are all preserved in the applicant's laboratory.

[0034] Main reagents: Bst DNA polymerase large fragment, New England Biolab; Betaine, Sigma; Tap DNA polymerase, deoxynucleoside triphosphate (dNTP Mixture), DL2000DNA Marker , 6×Loading buffer, TaKaRa Biosynthesis Company; DNAGreen, TIANDZ; SYBR Green I, Solarbio; Genomic DNA Rapid Extraction Kit (blood), Shanghai Sangon Biosynthesis Co., Ltd.

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Abstract

The invention discloses a method for detecting anaplasma phagocytophilum employing a loop-mediated isothermal amplification (LAMP) technology. The detection method comprises an LAMP primer design, building of an LAMP reaction system and adoption of an LAMP detection method. Specificity detection, sensitivity detection and fluorescent visual detection are adopted in the LAMP detection method. The anaplasma phagocytophilum 16SrRNA is taken as a target gene locus, and has the characteristics of being fast, sensitive, efficient, low in cost and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for detecting phagocytophilic aplasma by using a loop-mediated isothermal amplification technique (LAMP). Background technique [0002] Anaplasma phagocytophilum is an important zoonotic pathogen belonging to the genus Anaplasma in the family Anaplasmataceae of the order Rickettsiale. The pathogen is widely parasitic in granulocytes including humans, wild animals, domestic animals and rodents. Human granulocytic anaplasmosis (Human granulocytic anaplasmosis, HGA) is caused by A.phagocytophilum invading human peripheral blood neutrophils (Lj Zhang, Y Liu, et al. Nosocomial Transmission of Human Granulocytic Anaplasmosis in China [J]. The Journal of the American Medical Association, 2008, 300(19): 2263-2270). [0003] At present, the main detection methods of anplasma include etiological diagnosis, serological diagnosis and molecular biology diagnosis. [00...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/04C12Q1/6844C12Q2531/119
Inventor 宁长申王金鸿菅复春吕亚莉张龙现王荣军张艳韩琼琼张文静
Owner HENAN AGRICULTURAL UNIVERSITY
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