TY - JOUR
T1 - Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR
AU - van den Berg, R. J.
AU - Kuijper, E. J.
AU - van Coppenraet, L. E.S.Bruijnesteijn
AU - Claas, E. C.J.
N1 - Funding Information: This work was supported by a grant from the Foundation Microbiology Leiden. We thank K. Templeton for support in the development of the real-time PCR assay. This study was presented, in part, at the 43rd Interscience Conference of Antimicrobial Agents and Chemotherapy (Chicago, 2003), and the First International Clostridium difficile Symposium (Slovenia, 2004).
PY - 2006/2
Y1 - 2006/2
N2 - A real-time PCR assay for Clostridium difficile was developed, based on the tcdB gene, which detected all known toxinogenic reference strains (n = 45), within 30 serogroups and 24 toxinotypes. The analytical sensitivity was 1 × 103 CFU/mL, and the detection limit in faeces was 1 × 105 CFU/g. The optimal protocol for DNA extraction from faecal samples involved use of the Magna-Pure system with a Stool Transport and Recovery (STAR) buffer pre-treatment. In a 1-month prospective study of 85 patients with diarrhoea, the sensitivity, specificity and positive and negative predictive values of the assay were 100%, 94%, 55% and 100%, respectively, compared with the standard cell cytotoxicity assay.
AB - A real-time PCR assay for Clostridium difficile was developed, based on the tcdB gene, which detected all known toxinogenic reference strains (n = 45), within 30 serogroups and 24 toxinotypes. The analytical sensitivity was 1 × 103 CFU/mL, and the detection limit in faeces was 1 × 105 CFU/g. The optimal protocol for DNA extraction from faecal samples involved use of the Magna-Pure system with a Stool Transport and Recovery (STAR) buffer pre-treatment. In a 1-month prospective study of 85 patients with diarrhoea, the sensitivity, specificity and positive and negative predictive values of the assay were 100%, 94%, 55% and 100%, respectively, compared with the standard cell cytotoxicity assay.
KW - Assay
KW - Clostridium difficile
KW - Faecal samples
KW - Real-time PCR
KW - tcdB gene
UR - http://www.scopus.com/inward/record.url?scp=33644881953&partnerID=8YFLogxK
U2 - https://doi.org/10.1111/j.1469-0691.2005.01301.x
DO - https://doi.org/10.1111/j.1469-0691.2005.01301.x
M3 - Article
C2 - 16441459
SN - 1198-743X
VL - 12
SP - 184
EP - 186
JO - Clinical Microbiology and Infection
JF - Clinical Microbiology and Infection
IS - 2
ER -