Kit for simultaneously detecting drug-resistant loci and virulence genotypes of helicobacter pylori and metabolic genotypes of proton pump inhibitors

Abstract

The invention provides a kit for simultaneously detecting drug-resistant loci and virulence genotypes of helicobacter pylori and metabolic genotypes of proton pump inhibitors. Specifically, 16 drug-resistant gene locus variations of 6 common helicobacter pylori antibiotics, 5 loci of 2 virulence genes and 3 loci of 2 proton pump drug metabolism genes are detected on the basis of multiple PCR-time-of-flight mass spectrometry, and matched primers, probe combinations and kits are provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a kit for simultaneously detecting drug resistance sites of Helicobacter pylori, virulence genotyping and proton pump inhibitor drug metabolism genotyping. Background technique [0002] Helicobacter pylori (H. pylori) is a microaerophilic Gram-negative bacterium that colonizes gastric epithelial cells, infects about 50% to 70% of the world's population, and the infection rate in some developing countries is even as high as 80%. One of the most prevalent pathogens causing gastrointestinal disorders including peptic ulcer disease, gastric marginal zone / mucosa-associated lymphoid tissue (MALT) lymphoma, and gastric cancer. Helicobacter pylori has been classified as a class I carcinogen by the World Health Organization and the International Agency for Research on Cancer (IARC). According to statistics, more than 6% of cancers worldwide and about 90% of noncardia g...

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Problems solved by technology

Chinese patents CN109797203A "Helicobacter pylori detection system and detection kit and application" and CN111334592A "a nucleic acid composition for detecting Helicobacter pylori drug resistance gene and its kit and application" respectively disclose the Fluorescent quantitative PCR amplification system for identification, typing and antibiotic resistance. However, due to the technical limitations of the current fluorescent quantitative PCR method, one detection reaction can only realize the simultaneous detection of several sites at most, and the detection throughput is limited. When It will be limited when more gene mutation points need to be detected

On the other hand, the conventional fluorescent quantitative PCR method may not be able to meet the needs of rapid detection of large samples

Chinese patent CN105368825A "Helicobacter pylori antibiotic resistance analysis kit and drug resistance detection method" and CN105506160A "Helicobacter pylori quantitative and virulence multiple gene detection system and its kit and application" both disclose a method based on multiplex PCR - Capillary electrophoresis analysis of antibiotic resistance of Helicobacter pylori commonly used in the same reaction system, but capillary electrophoresis can only be typed according to fragment size, and has high uniformity and specificity for multiplex PCR amplification Require

However, due to the complex and expensive equipment and process of chip detection and high-throughput sequencing (next-generation and third-generation sequencing), the detection cost is high and time-consuming, so it is mainly used for the correlation research between genotype and phenotype drug resistance. On the one hand, it is rarely used in the clinical molecular detection of Helicobacter pylori

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