Detection of toxigenic Clostridium difficile in stool specimens by the polymerase chain reaction.
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.
Kato
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Nosocomial Pathogens Laboratory Branch, Centers for Disease Control, Atlanta, Georgia.
Ou
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Kato
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Bartley
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SL
Luo
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CC
Killgore
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GE
Ueno
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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
D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; D009838:Oligodeoxyribonucleotides; C041348:tcdA protein, Clostridium difficile