Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR

R. J. van den Berg; E. J. Kuijper; L. E.S.Bruijnesteijn van Coppenraet; E. C.J. Claas

doi:10.1111/j.1469-0691.2005.01301.x

Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR

R. J. van den Berg
, E. J. Kuijper^*
, L. E.S.Bruijnesteijn van Coppenraet
, E. C.J. Claas

^*Corresponding author for this work

Leiden University

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

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.

Original language	English
Pages (from-to)	184-186
Number of pages	3
Journal	Clinical microbiology and infection
Volume	12
Issue number	2
DOIs	https://doi.org/10.1111/j.1469-0691.2005.01301.x
Publication status	Published - Feb 2006

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Assay
Clostridium difficile
Faecal samples
Real-time PCR
tcdB gene

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10.1111/j.1469-0691.2005.01301.x

Cite this

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title = "Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR",

abstract = "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.",

keywords = "Assay, Clostridium difficile, Faecal samples, Real-time PCR, tcdB gene",

author = "\{van den Berg\}, \{R. J.\} and Kuijper, \{E. J.\} and \{van Coppenraet\}, \{L. E.S.Bruijnesteijn\} and Claas, \{E. C.J.\}",

note = "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).",

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doi = "10.1111/j.1469-0691.2005.01301.x",

language = "English",

volume = "12",

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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).

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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 - https://www.scopus.com/pages/publications/33644881953

U2 - 10.1111/j.1469-0691.2005.01301.x

DO - 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 -

Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR

Abstract

UN SDGs

Keywords

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