title: Detection of Clostridium difficile toxin A/B genes by multiplex real-time PCR for the diagnosis of C. difficile infection.
abstract: Toxigenic Clostridium difficile culture is considered to be the standard diagnostic method for the detection of C. difficile infection (CDI). Culture methods are time-consuming and although enzyme immunoassay is rapid and easy to use, it has low sensitivity. In the present study, the AdvanSure CD real-time (RT)-PCR kit (LG Life Sciences) was evaluated for its ability to detect C. difficile toxin A (tcdA) and B (tcdB) genes, simultaneously. A total of 127 fresh diarrhoeal stool specimens, submitted to the clinical microbiology laboratory for C. difficile culture, were tested. C. difficile toxins and toxin genes were detected with a VIDAS C. difficile A&B (VIDAS-CDAB) enzyme-linked fluorescent immunoassay (ELFA) and the AdvanSure RT-PCR kit, respectively, according to the manufacturers' instructions. Their performance was compared with a standard toxigenic culture method as a reference. The sensitivity, specificity and positive and negative predictive values using the AdvanSure RT-PCR kit were 100 %, 98.3 %, 84.6 % and 100 %, respectively, while those of the VIDAS-CDAB system were 63.6 %, 100 %, 100 % and 96.6 %, respectively. Four tcdA(+)/tcdB(+) strains of C. difficile were detected with the AdvanSure RT-PCR kit, which offers comparable sensitivity and specificity to the reference method with a turnaround time of ~3 hours.
authors.affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Republic of Korea.
authors.forename: Heejung
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Republic of Korea.
authors.forename: Seok Hoon
authors.initials: SH
authors.lastname: Jeong
authors.affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Republic of Korea.
authors.forename: Myungsook
authors.initials: M
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Republic of Korea.
authors.forename: Yangsoon
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Republic of Korea.
authors.forename: Kyungwon
authors.initials: K
authors.lastname: Lee
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods; D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification*; D003015:Clostridium Infections / Q000175:diagnosis* / Q000382:microbiology; D004768:Enterotoxins / Q000032:analysis* / Q000235:genetics; D005243:Feces / Q000382:microbiology; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; D011933:Reagent Kits, Diagnostic; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile