title: Diagnosis of Clostridium difficile infection using real-time PCR.
abstract: Clostridium difficile is known to cause antibiotic-associated diarrhea and pseudomembranous colitis. Toxinogenic strains of the bacterium produce toxins A (TcdA) and B (TcdB), which are associated with the pathogenicity. The standard methods for diagnosis of C. difficile infection include the cell cytotoxicity assay and the culture of a toxinogenic strain. Due to the long turnaround time of these methods, more rapid methods are preferred. Enzyme immunoassays are fast, but lack sensitivity. Therefore, real-time PCR methods have been developed. The real-time PCR described in this chapter detects tcdB, the gene coding for toxin B. Since toxin A-negative, toxin B-positive strains have been reported to cause disease as well, these strains can also be detected by this method which uses an automated STAR-MagnaPure method for the optimum isolation of DNA from feces. An internal control is included as well to control for inhibition of the PCR method.
authors.affiliation: Department of Medical Microbiology, Centre of Infectious Diseases, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Renate Johanna
authors.initials: RJ
authors.lastname: van den Berg
authors.forename: Dennis
authors.initials: D
authors.lastname: Bakker
authors.forename: Ed J
authors.initials: EJ
authors.lastname: Kuijper
mesh_terms: D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification*; D004247:DNA / Q000302:isolation & purification; D004761:Enterocolitis, Pseudomembranous / Q000175:diagnosis*; D005243:Feces / Q000737:chemistry / Q000382:microbiology; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012984:Software
chemical_list: D004247:DNA