title: Detection of toxigenic Clostridium difficile in stool specimens by the polymerase chain reaction.
abstract: Polymerase chain reaction (PCR) amplification of a segment of the toxin A gene was used to detect toxigenic Clostridium difficile directly from stool specimens of patients with antibiotic-associated diarrhea. Although PCR-inhibitory substances were recognized in DNA prepared from stool specimens, the inhibitory substances were eliminated by using an ion-exchange column after phenol-chloroform extraction. Eventually, 39 stool specimens were evaluated by PCR. PCR results for detection of toxigenic C. difficile were in complete agreement with cell culture assay results; all 12 PCR-positive stool specimens were positive by cytotoxin assay, and all 27 PCR-negative specimens were negative by cytotoxin assay. Toxigenic C. difficile was cultured from all PCR-positive specimens. These results suggest that PCR amplification may be an effective method for laboratory diagnosis of C. difficile-associated diarrhea and colitis.
authors.affiliation: Nosocomial Pathogens Laboratory Branch, Centers for Disease Control, Atlanta, Georgia.
authors.forename: N
authors.initials: N
authors.lastname: Kato
authors.forename: C Y
authors.initials: CY
authors.lastname: Ou
authors.forename: H
authors.initials: H
authors.lastname: Kato
authors.forename: S L
authors.initials: SL
authors.lastname: Bartley
authors.forename: C C
authors.initials: CC
authors.lastname: Luo
authors.forename: G E
authors.initials: GE
authors.lastname: Killgore
authors.forename: K
authors.initials: K
authors.lastname: Ueno
mesh_terms: D001427:Bacterial Toxins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry; D004761:Enterocolitis, Pseudomembranous / Q000382:microbiology*; D004768:Enterotoxins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D005243:Feces / Q000382:microbiology*; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; D009838:Oligodeoxyribonucleotides; C041348:tcdA protein, Clostridium difficile