title: Detection of virulence genes of Clostridium difficile by multiplex PCR.
abstract: Clostridium difficile strains belonging to the PCR ribotype 027, pulse-field gel electrophoresis (PFGE) type NAP1, toxinotype III and restriction endonuclease analysis group BI harbouring mutations in the tcdC gene and possessing binary toxin components A and B have been described to cause epidemics with increased morbidity and mortality. In the present study we developed a conventional multiplex PCR designed to detect selected virulence associated markers of the hypervirulent C. difficile PCR ribotype 027. The multiplex PCR assay detected the major toxins A and B, binary toxin components A and B as well as a possible deletion in the tcdC gene: a characteristic pattern of amplification products for the PCR ribotype 027 strains was detected. This rather simple method was specific for the screening of this hypervirulent C. difficile strain. The correlation between the multiplex PCR and PCR ribotyping methods was excellent. The sensitivity and specificity were 100% in our epidemiological situation. In conclusion, this multiplex PCR was found useful in the preliminary screening for the hypervirulent C. difficile PCR ribotype 027.
authors: Division of Clinical Microbiology, HUSLAB, Helsinki University Central Hospital, Helsinki, Finland.
authors: Jenni
authors: J
authors: Antikainen
authors: Tanja
authors: T
authors: Pasanen
authors: Sointu
authors: S
authors: Mero
authors: Eveliina
authors: E
authors: Tarkka
authors: Juha
authors: J
authors: Kirveskari
authors: Saara
authors: S
authors: Kotila
authors: Silja
authors: S
authors: Mentula
authors: Eija
authors: E
authors: Könönen
authors: Anni-Riitta
authors: AR
authors: Virolainen-Julkunen
authors: Martti
authors: M
authors: Vaara
authors: Päivi
authors: P
authors: Tissari
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics*; D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity*; D004269:DNA, Bacterial / Q000032:analysis; D004761:Enterocolitis, Pseudomembranous / Q000175:diagnosis* / Q000382:microbiology; D004768:Enterotoxins / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012097:Repressor Proteins / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; D012097:Repressor Proteins; C105615:TcdC protein, Clostridium difficile; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile