Detection of peptostreptococcus micros DNA in clinical samples by PCR.
Peptostreptococcus micros is a gram-positive anaerobic coccus which, although considered to be a natural commensal of the human oral cavity, is associated with periodontal, endodontal and peritonsillar infections. Identification of the organism has to date relied upon conventional culture methods and biochemical analyses. The purpose of this study was to develop a PCR method for rapid and specific identification of this organism in clinical samples. A pair of primers was selected, each of which was specific at the 3' end for P. micros DNA; they were used in the PCR assay, resulting in a 1074-bp product. The primers were shown to be specific for P. micros DNA as no PCR products were obtained when genomic DNA extracts from a wide range of other Peptostreptococcus species and other oral bacteria were used as templates. The PCR assay was then applied to the identification of P. micros DNA in subgingival plaque samples from adult periodontitis patients and pus samples from subjects with acute dento-alveolar abscesses. Confirmation of specific amplification of P. micros DNA was obtained by digestion of PCR products with the restriction endonuclease RsaI, which gives a unique restriction profile for P. micros, and DNA sequencing. Sixty-eight subgingival plaque samples from 18 patients were analysed, of which 19 (28%) were positive for P. micros DNA; the proportion of patients carrying P. micros DNA in at least one sampled site was 11 (61%) of 18. Twenty (71%) of 28 pus samples analysed by PCR contained P. micros DNA. These results confirm that P. micros may be involved in the aetiology of acute dentoalveolar abscesses and adult periodontitis. The PCR assay provides a more rapid and reliable alternative to conventional methods for identification of P. micros in clinical samples.
Infection Research Group, University of Glasgow Dental School, 378 Sauchiehall Street, Glasgow G2 3JZ.
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Riggio
Infection Research Group, University of Glasgow Dental School, 378 Sauchiehall Street, Glasgow G2 3JZ.
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Lennon
Infection Research Group, University of Glasgow Dental School, 378 Sauchiehall Street, Glasgow G2 3JZ.
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Smith
D000328:Adult; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D008875:Middle Aged; D010462:Peptostreptococcus / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial