Laboratory diagnosis of toxigenic Clostridium difficile by polymerase chain reaction: Presence of toxin genes and their stable expression in toxigenic isolates from Japanese individuals.
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概要
- 論文の詳細を見る
Clostridium difficile causes pseudomembranous colitis and antibiotic-associated diarrhea. The definitive diagnosis of C. difficile infection is finally accomplished by the isolation of toxigenic C. difficile. However, only a small number of Japanese clinical laboratories are able to reach a definitive diagnosis of C. difficile infection, probably because simple reliable assays for toxins in the isolates are not available. In this study, we examined the compatibility of a polymerase chain reaction (PCR) assay and tissue culture assay to identify toxigenic C. difficile, in toxigenic and nontoxigenic C. difficile isolates from Japanese patients and healthy carriers. The specificity of PCR primers was demonstrated by restriction endonuclease digestion and seminested PCR in C. difficile VPI 10463 strain. No PCR product was amplified in the eight other clostridial species used to check the specifiecty of the PCR assay. The detection limit was 103 cells. Both toxin A and toxin B genes (the genes encoding the major virulence factors of C. difficile) were detected in 58 toxigenic C. difficile isolates, which showed a wide range of cytotoxic activity in tissue culture assays. Neither of the toxin genes was carried by 40 nontoxigenic strains of C. difficile. The results of this study strongly suggest that a definitive diagnosis of C. difficile infection can be accomplished by PCR detection of the toxin genes rather than by tissue culture assay of isolates.
- Springer Verlagの論文
- 1999-02-01
著者
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Nakamura Shinichi
Department of Cardiovascular Medicine, Kumamoto Central Hospital
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KARASAWA Tadahiro
Department of Laboratory Sciences, School of Health Sciences, Kanazawa University
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Karasawa T
Kanazawa Univ. Ishikawa Jpn
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Karasawa T
Kanazawa Univ. Ishikawa
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Karasawa Tadahiro
Department Of Clinical Laboratory Science Division Of Health Sciences Kanazawa University Graduate S
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HAYASHI YASUHIRO
Department of Materials Science, Wakayama National College of Technology
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Nakamura Shinichi
Department Of Microbiology School Of Medicine Kanazawa University
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WANG Xing
Department of Chemistry, Graduate School of Science, Tohoku University and CREST, Japan Science and
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NOJIRI TAKASHI
Department of Bacteriology, School of Medicine, Kanazawa University
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MAEGAWA TSUNEO
Department of Bacteriology, School of Medicine, Kanazawa University
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YAMAKAWA KIYOTAKA
Department of Bacteriology, School of Medicine, Kanazawa University
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Maegawa Tsuneo
Department Of Bacteriology School Of Medicine Kanazawa University
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Yamakawa Kiyotaka
Department Of Bacteriology School Of Medicine Kanazawa University
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Yamakawa Kiyotaka
Department Of Bacteriology Kanazawa University School Of Medicine
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Hayashi Yasuhiro
Department Of Material Science Wakayama National College Of Technology
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Wang Xing
Department Of Bacteriology School Of Medicine Kanazawa University
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Hayashi Yasuhiro
Department Of Bacteriology School Of Medicine Kanazawa University
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Nakamura Shinichi
Department Of Anesthesiology Saitama Medical School
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Wang XingMin
Department of Bacteriology, School of Medicine, Kanazawa University
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