Detection of Bacteroides fragilis in clinical specimens by polymerase chain reaction amplification of the neuraminidase gene |
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Authors: | Ravi Jotwani Naoki Kato Haru Kato Kunitomo Watanabe Kazue Ueno |
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Affiliation: | (1) Institute of Anaerobic Bacteriology, Gifu University School of Medicine, 40 Tsukasa-machi, 500 Gifu, Japan;(2) Present address: Department of Microbiology, National Institute of Immunology, Aruna Asaf Ali Marg, 110067 New Delhi, India;(3) Present address: Gifu College of Medical Technology, 795-1 Nagamine, 501-32 Seki, Japan |
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Abstract: | The polymerase chain reaction (PCR) was used in an attempt to detect Bacteroides fragilis by amplifying a segment of the gene encoding B. fragilis neuraminidase. Forty-five reference strains representing 45 species and 113 clinical isolates were tested. Only B. fragilis was PCR positive, except for Bacteroides merdae ATCC 43184, which gave a band by ethidium bromide staining that showed no signal by Southern hybridization. Using a protocol that employed DNA extraction by Sepa Gene kit and a highly sensitive digoxigenin-chemiluminescence detection system, detection of B. fragilis by PCR was in complete agreement with culture results for 44 clinical specimens from which a wide range of aerobic and anaerobic organisms and fungi were recovered. |
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