Establishment of methodology for carrying out joint detection on bacterial genus genes and toxin genes of clostridium difficile by using TaqMan-MGB probe real-time fluorescent quantitative PCR (polymerase chain reaction) technology

Abstract

A TaqMan/MGB probe PCR (polymerase chain reaction) technology can carry out bacterial genus identification on clostridium difficile in fecal genomes, simultaneously detect the carrying situation of toxin genes, and can judge whether a toxin A has deletion. A fecal specimen is not required to be purely cultured, and the strain identification and the toxin detection are completed in a reaction system. The method has the characteristics of simple operation, good repeatability, high flux detection specimens, short report time, and the like, and is applicable to the screening of pathogenesis of patients with diarrhea caused by clinically unexplained causes. The invention solves the technical problems that fecal specimens can be subjected to strain identification and strain toxin carrying situation screening simultaneously without being purely cultured, and a DNA (deoxyribonucleic acid) detection method for fecal genomes, which is high in sensibility, is provided. Because traditional anaerobic culture is uneasy to perform, and an enzyme immunoassay has methodology defects, according to the invention, the diagnosis rate of clostridium difficile associated diarrhea is significantly increased. Meanwhile, the invention also can be applied to the monitoring of drug used in the process of treating the diseases.

Description

technical field [0001] The patent of the present invention relates to a new detection method of Clostridium difficile gene and toxin gene, which specifically uses TaqMan / MGB probe technology to quickly perform fecal genomic DNA amplification and fluorescent signal detection through the Real-TimePCR analysis system, and simultaneously complete the difficult Identification of Clostridium, detection of A and B toxin genes, and identification of whether A toxin is missing. Background technique [0002] Clostridium difficile has always been the main pathogen causing iatrogenic diarrhea. Due to the extensive use of broad-spectrum antibiotics and the highly virulent strain 027 / NAP1 / BI (nucleic acid type is 027, pulse-field gel electrophoresis type is NAP1, restricted With the emergence and prevalence of endonuclease type BI), the epidemic outbreak of CDI in the world, especially in Europe and North America, has increased rapidly, the number of severe cases, recurrence rate and fata...

AI Technical Summary

Problems solved by technology

It can be seen that A-B+ strains in China have the characteristics of high incidence rate, strong concealment and high drug resistance, and are more harmful to clinical departments.