Oligonucleotide primer for detecting common pathogenic bacteria by adopting fluorescent quantitation PCR (Rich Client Platform) technology, method thereof for detecting common pathogenic bacteria and application thereof

Abstract

The invention discloses an oligonucleotide primer for detecting common pathogenic bacteria by adopting a fluorescent quantitation PCR (Rich Client Platform) technology, a method thereof for detecting common pathogenic bacteria and the application thereof. The method comprises the following steps of: providing 10 pairs of specific oligonucleotide primer sequences at annealing temperature of 50-60 DEG C without differing 5 DEG C; and simultaneously, quickly, accurately and effectively identifying and quantificationally detecting various pathogenic bacteria at the same time. A detection range comprises bacillus cereus, enterobacter sakazakii, vibrio parahaemolyticus, enterohemorrhagic escherichia coli O157, salmonella, Listeria monocytogenes, Shigella, campylobacter jejuni, pseudomonas aeruginosa, klebsiella pneumoniae, and the like. The invention also can be used for the fields of disease diagnosis, environmental monitoring, water-quality and food supervision and detection, food poisoning pathogenicbacteria detection, bacteriological classification, epidemiological investigation, biological agent detection, and the like, is convenient, quick, accurate and effective and has wide application range.

Description

technical field [0001] The invention relates to the technical field of pathogenic bacteria detection, in particular to a convenient, fast, accurate, efficient and wide application oligonucleotide primer for detecting common pathogenic bacteria using fluorescence quantitative PCR technology and detecting common pathogenic bacteria Methods and uses of bacteria. Background technique [0002] As we all know, the detection of pathogenic bacteria has important applications in the fields of disease diagnosis, environmental monitoring, food poisoning detection, import and export food quarantine, scientific research and other fields. Traditional pathogenic bacteria testing methods still remain at the level of isolation and culture, morphological observation, biochemical identification, and serological typing, which not only takes a long time, but also cannot effectively detect pathogenic bacteria that are difficult to culture, and can only be used for limited species one by one. det...

AI Technical Summary

Problems solved by technology

[0003] Conventional PCR can realize the rapid detection of microorganisms, especially pathogenic bacteria, but it cannot quantify the pathogenic bacteria to be detected, and only one kind of pathogenic bacteria can be detected in one experiment, and the efficiency is low; quantitative PCR can be carried out for the pathogenic bacteria to be detected Quantitative detection, but one experiment can only detect one pathogenic bacteria to be tested, and cannot achieve high-throughput (simultaneous) detection of pathogenic bacteria; although multiple PCR and gene chips have the ability to detect several pathogenic bacteria at one time, However, the biggest shortcoming of these two methods is that the pathogenic bacteria to be detected cannot be quantified. In addition, multiplex PCR has high requirements for primers, and there is an electrophoresis stage, which is prone to cross-infection and false positives; Needle