Dual fluorescence quantitative PCR (polymerase chain reaction) detection method and detection kit for clostridium difficile enterotoxin A and B

Abstract

The invention discloses a dual fluorescence quantitative PCR (polymerase chain reaction) detection method and a detection kit for clostridium difficile enterotoxin A and B. The method comprises the steps that 1) the DNA (deoxyribonucleic acid) of a sample to be detected is extracted; 2) the DNA (deoxyribonucleic acid) of the sample to be detected is used as a template and is subjected to fluorescence quantitative PCR reaction; 3) the fluorescence of each cyclic product in the PCR reaction is subjected to fluorescence detection, and whether the sample to be detected contains clostridium difficile enterotoxin A and B or not is judged according to the lowest Ct value and the highest fluorescence value in the fluorescence detection. The invention also discloses a specific primer, a fluorescence probe and the detection kit. The dual fluorescence quantitative PCR detection method is simple, convenient and quick to operate, can detect the clostridium difficile enterotoxin A and B simultaneously, is high in detection sensitivity and specificity and can be applicable to the laboratory emergency detection of outburst epidemic caused by clostridium difficile.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a detection method and a detection kit for Clostridium difficile enterotoxin A and B. Background technique [0002] Clostridium difficile (Clostridium difficile) is a Gram-positive anaerobic bacillus with a spore structure. It is recognized as the most important pathogen of nosocomial infection and antibiotic-associated diarrhea. It can cause pseudomembranous colitis and even lethal by releasing cytotoxin. Clostridium difficile can release a variety of toxins, among which toxins A and B are the main ones, which are the main causes of diarrhea and intestinal inflammation. [0003] Clostridium difficile laboratory diagnosis methods mainly rely on traditional anaerobic culture, cytotoxicity test, bacteria and toxin antigen detection. Among them, bacterial anaerobic culture and cytotoxin neutralization experiments are currently the "gold standard" for laboratory detection of Clostridium d...

AI Technical Summary

Problems solved by technology

Glutamate dehydrogenase (GDH) is an antigenic enzyme protein abundantly expressed on the surface of Clostridium difficile, but immunoenzyme tests cannot distinguish toxin-producing strains of Clostridium difficile from non-toxigenic strains

In addition, the current commercialized immunological C. difficile toxin diagnostic products can only detect toxin A, which leads to the missed diagnosis of many patients caused by clinical A-B+ C. difficile, and makes serious infections and outbreaks caused by A-B+ C. increase

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