Tags

Type your tag names separated by a space and hit enter

New multiplex PCR method for the detection of Clostridium difficile toxin A (tcdA) and toxin B (tcdB) and the binary toxin (cdtA/cdtB) genes applied to a Danish strain collection.
Clin Microbiol Infect. 2008 Nov; 14(11):1057-64.CM

Abstract

Isolates of Clostridium difficile from 159 hospitalized Danish patients (2005) were analysed by a new 5-plex PCR method targeting the toxin genes tcdA, tcdB, cdtA and cdtB, and 16S rDNA as an internal positive control. Additionally, the toxin-regulating gene tcdC was partially sequenced by a new sequencing-based method that revealed genetic changes that may render the gene product inactive. Finally tcdA was analysed using a previously published method for the detection of internal deletions. The 5-plex PCR revealed four different toxin gene profiles: 36 tcdA+, tcdB+, cdtA+/cdtB+; one tcdA+, tcdB-, cdtA+/cdtB+; 98 tcdA+, tcdB+, cdtA-/cdtB-; and 24 non-toxigenic tcdA-, tcdB-, cdtA-/cdtB-. Deletion studies revealed that 26 strains contained a c. 700-bp deletion in tcdA, and 39 strains contained at least one possible inactivation feature in tcdC. The prevalence of the binary toxin genes was 23%. All strains with the tcdA+, tcdB+, cdtA+/cdtB+ profile were investigated by PCR ribotyping, and this revealed eight different ribotypes, none of which were 027. The 5-plex PCR method offers a one-step, rapid and specific screening method for C. difficile toxin genes. This toxin gene profiling, together with deletion studies in tcdA and tcdC, may allow an evaluation of the pathogenic potential of C. difficile.

Authors+Show Affiliations

Department of Bacteriology, Mycology and Parasitology, TheNational Reference Laboratory for Enteropathogenic Bacteria, Unit of Gastrointestinal Infection, Statens Serum Institut, Copenhagen, Denmark. spn@ssi.dkNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

19040478

Citation

Persson, S, et al. "New Multiplex PCR Method for the Detection of Clostridium Difficile Toxin a (tcdA) and Toxin B (tcdB) and the Binary Toxin (cdtA/cdtB) Genes Applied to a Danish Strain Collection." Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, vol. 14, no. 11, 2008, pp. 1057-64.
Persson S, Torpdahl M, Olsen KE. New multiplex PCR method for the detection of Clostridium difficile toxin A (tcdA) and toxin B (tcdB) and the binary toxin (cdtA/cdtB) genes applied to a Danish strain collection. Clin Microbiol Infect. 2008;14(11):1057-64.
Persson, S., Torpdahl, M., & Olsen, K. E. (2008). New multiplex PCR method for the detection of Clostridium difficile toxin A (tcdA) and toxin B (tcdB) and the binary toxin (cdtA/cdtB) genes applied to a Danish strain collection. Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, 14(11), 1057-64. https://doi.org/10.1111/j.1469-0691.2008.02092.x
Persson S, Torpdahl M, Olsen KE. New Multiplex PCR Method for the Detection of Clostridium Difficile Toxin a (tcdA) and Toxin B (tcdB) and the Binary Toxin (cdtA/cdtB) Genes Applied to a Danish Strain Collection. Clin Microbiol Infect. 2008;14(11):1057-64. PubMed PMID: 19040478.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - New multiplex PCR method for the detection of Clostridium difficile toxin A (tcdA) and toxin B (tcdB) and the binary toxin (cdtA/cdtB) genes applied to a Danish strain collection. AU - Persson,S, AU - Torpdahl,M, AU - Olsen,K E P, PY - 2008/12/2/pubmed PY - 2009/1/3/medline PY - 2008/12/2/entrez SP - 1057 EP - 64 JF - Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases JO - Clin Microbiol Infect VL - 14 IS - 11 N2 - Isolates of Clostridium difficile from 159 hospitalized Danish patients (2005) were analysed by a new 5-plex PCR method targeting the toxin genes tcdA, tcdB, cdtA and cdtB, and 16S rDNA as an internal positive control. Additionally, the toxin-regulating gene tcdC was partially sequenced by a new sequencing-based method that revealed genetic changes that may render the gene product inactive. Finally tcdA was analysed using a previously published method for the detection of internal deletions. The 5-plex PCR revealed four different toxin gene profiles: 36 tcdA+, tcdB+, cdtA+/cdtB+; one tcdA+, tcdB-, cdtA+/cdtB+; 98 tcdA+, tcdB+, cdtA-/cdtB-; and 24 non-toxigenic tcdA-, tcdB-, cdtA-/cdtB-. Deletion studies revealed that 26 strains contained a c. 700-bp deletion in tcdA, and 39 strains contained at least one possible inactivation feature in tcdC. The prevalence of the binary toxin genes was 23%. All strains with the tcdA+, tcdB+, cdtA+/cdtB+ profile were investigated by PCR ribotyping, and this revealed eight different ribotypes, none of which were 027. The 5-plex PCR method offers a one-step, rapid and specific screening method for C. difficile toxin genes. This toxin gene profiling, together with deletion studies in tcdA and tcdC, may allow an evaluation of the pathogenic potential of C. difficile. SN - 1469-0691 UR - https://www.unboundmedicine.com/medline/citation/19040478/New_multiplex_PCR_method_for_the_detection_of_Clostridium_difficile_toxin_A__tcdA__and_toxin_B__tcdB__and_the_binary_toxin__cdtA/cdtB__genes_applied_to_a_Danish_strain_collection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1198-743X(14)61817-6 DB - PRIME DP - Unbound Medicine ER -