Diagnosis of Clostridium difficile infection using real-time PCR.

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.

Citation

Renate Johanna van den Berg, Dennis Bakker, Ed J Kuijper. Diagnosis of Clostridium difficile infection using real-time PCR. Methods in molecular biology (Clifton, N.J.). 2013;943:247-56

Mesh Tags

Substances

PMID: 23104294

search → result