A loop-mediated isothermal amplification detection method and a kit of a high-virulence strain of Helicobacter pylori

A Helicobacter pylori, loop-mediated isothermal technology, applied in the field of medical molecular biology diagnosis, can solve problems such as lack of suitable primers, and achieve the effects of shortening detection time, avoiding pollution and random errors, and high specificity

Pending Publication Date: 2019-01-11
JINAN CENTER HOSPITAL
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Problems solved by technology

In recent years, this technology has been gradually applied to the detection of pathogenic microorganisms, but so far this method has not been applied to the detection of highly virulent Helicobacter pylori, and there is no suitable and efficient primer for the detection of LAMP in this field

Method used

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  • A loop-mediated isothermal amplification detection method and a kit of a high-virulence strain of Helicobacter pylori
  • A loop-mediated isothermal amplification detection method and a kit of a high-virulence strain of Helicobacter pylori
  • A loop-mediated isothermal amplification detection method and a kit of a high-virulence strain of Helicobacter pylori

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Experimental program
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Embodiment

[0072] 1. Materials and methods

[0073] 1.1 Experimental strains and clinical samples

[0074] A total of 10 strains of bacteria were used in this study, Helicobacter pylori NCTC11637 (as a positive control for the determination of specificity and sensitivity), Streptococcus pneumoniae ATCC6305, Streptococcus agalactiae ATCC 9925, Enterococcus faecalis ATCC29212, Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Klebsiella pneumoniae ATCC700603, Pseudomonas aeruginosa ATCC27853, Streptococcus pyogenes ATCC19615 were purchased from the Institute of Microbiology, Chinese Academy of Sciences, and the clinical isolates of Helicobacter pylori were provided by the strain bank of our laboratory.

[0075] 188 clinical samples, including gastric mucosal tissue, gastric juice, intestinal juice, saliva, dental plaque, feces, etc., were collected from first-time patients of different ages and genders in the Department of Gastroenterology.

[0076] 1.2 DNA extraction

[0077]...

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Abstract

The invention provides a loop-mediated isothermal amplification detection method and a kit of a high-virulence strain of Helicobacter pylori. Five LAMP primers were designed for the nucleic acid sequence of Helicobacter pylori CagA gene, including external primers F3, B3; The internal primers FIP, BIP and loop primer LB ensured the high specificity of LAMP amplification. This technique is specificfor the detection of high virulence Helicobacter pylori strains, and has the characteristics of high speed, high efficiency, simple instrument requirements, simple operation, low cost, and easy to bepopularized and used in the grass roots. At the same time, the LAMP detection prim of the high-virulence strain of helicobacter pylori is designed and screen for the first time, and a LAMP detectionmethod of the high-virulence strain of helicobacter pylori is established, which fills a gap in the detection field at home and abroad.

Description

technical field [0001] The invention belongs to the technical field of medical molecular biology diagnosis, and in particular relates to a detection method and kit for a loop-mediated isothermal amplification of a highly virulent strain of Helicobacter pylori. Background technique [0002] The statements in this section merely provide background information related to the present disclosure and do not necessarily constitute prior art. [0003] Helicobacter pylori (Helicobacter pylori, HP) is a microaerophilic, spiral-shaped Gram-negative bacillus under the microscope. It is the only microbial species known to survive in human gastric juice. Epidemiological surveys show that the infection rate of HP in the general population is as high as 50% in the world, and it even reaches 70% to 90% in developing countries. There are also differences in the distribution of infections. The more backward the economy and the lower the education level, the higher the infection rate. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2563/107
Inventor 汪运山曹翠明杨炜华裴凤艳
Owner JINAN CENTER HOSPITAL
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