Quadruple fluorescent quantitative polymerase chain reaction (PCR) detection kit for diarrheagenic escherichia coli

Abstract

The invention provides a quadruple fluorescent quantitative polymerase chain reaction (PCR) detection kit for diarrheagenic Escherichia coli. The quadruple fluorescent quantitative PCR detection kit consists of fluorescent quantitative PCR reaction liquid, a standard substance O157, a standard substance H7, a standard substance O104, a standard substance H4 and a reference substance, wherein the fluorescent quantitative PCR reaction liquid comprises PCR reaction buffer solution (which contains a mixture of magnesium chloride and deoxyribose nucleotide triphosphate, heat-resisting thermus aquaticus deoxyribonucleic acid (TaqDNA) polymerase and the like), special upstream and downstream primers and a probe. In the quadruple fluorescent quantitative PCR detection kit, O157:H7 and O 104:H4 type bacteria of the diarrheagenic Escherichia coli can be detected from a sample; and compared with the conventional ordinary PCR method and single fluorescent quantitative PCR method, the quadruple fluorescent quantitative PCR detection kit has the advantages of simplicity, convenience, quickness and accuracy; and the detected bacteria are quantitated accurately in real time, and the diagnosis of suspected patients infected with the bacteria clinically is clarified early, so that the treatment scheme is formulated in time to lower the death rate.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a nucleic acid detection method for simultaneously detecting diarrhea-causing Escherichia coli O157:H7 and O104:H4 serotypes in stool samples of patients with gastroenteritis and diarrhea by quadruple real-time fluorescent quantitative PCR in the same reaction tube , can be applied to laboratory emergency detection of outbreaks caused by diarrhea-causing Escherichia coli. Background technique [0002] E. coli that causes diarrhea is called diarrhea-causing E. coli. According to virulence factors, pathogenic mechanisms, and epidemiological characteristics, diarrhea-causing E. coli is usually divided into five categories, namely enteropathogenic E. coli (EPEC) , enterotoxigenic Escherichia coli (ETEC), enteroinvasive Escherichia coli (EIEC), enteroaggregative Escherichia coli (EAEC) and enterohemorrhagic Escherichia coli (EHEC). Following EHEC infection, patients often present w...

AI Technical Summary

Problems solved by technology

[0003] In view of the increasing importance of diarrhea-causing Escherichia coli in public health, the detection of various diarrheal bacteria in the surveillance of gastroenteritis and food poisoning is not only time-consuming and laborious but also a waste of clinical specimens and reagents

Routine detection is mainly based on traditional culture methods, which are not only cumbersome to operate, time-consuming, but also easy to contaminate

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