TY - CHAP
T1 - Diagnosis of clostridium difficile infection using real-time PCR
AU - Van Den Berg, Renate Johanna
AU - Bakker, Dennis
AU - Kuijper, Ed J.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - Clostridium difficile
KW - DNA isolation from feces
KW - Pathogenicity locus
KW - Real-time PCR
KW - Toxin B
UR - http://www.scopus.com/inward/record.url?scp=84870505725&partnerID=8YFLogxK
U2 - https://doi.org/10.1007/978-1-60327-353-4_16
DO - https://doi.org/10.1007/978-1-60327-353-4_16
M3 - Chapter
C2 - 23104294
SN - 9781603273527
T3 - Methods in Molecular Biology
SP - 247
EP - 256
BT - PCR Detection of Microbial Pathogens
A2 - Wilks, Mark
ER -