Development of a novel PCR assay for detection of Prevotella oris in clinical specimens.
Prevotella oris is a nonpigmented, Gram-negative, anaerobic bacterium that has been associated with several serious oral and systemic infections. Prevotella oris has been identified in clinical specimens by bacterial culture and biochemical tests, which are generally unreliable. The aim of this study was to develop a PCR assay for the direct detection of P. oris in clinical specimens. PCR primers specific for P. oris were identified by alignment of bacterial 16S rRNA genes from closely related species and selection of PCR primers specific for P. oris at their 3' ends. Amplification of a 1110-bp product indicated PCR positivity for P. oris. The primers were shown to be specific for P. oris DNA, because no PCR products were obtained when DNA from other oral bacteria, including closely related Prevotella species, were used as test species, and this was confirmed by digestion of PCR products with RsaI and MnlI. Prevotella oris DNA was detected in 17 (36.2%) of 47 pus samples from subjects with dentoalveolar abscesses and in all three pus samples from subjects with spreading odontogenic infections. This PCR assay provides a sensitive, specific and reliable method for identifying P. oris in clinical specimens.
Riggio
Marcello Pasquale
MP
Infection and Immunity Section, University of Glasgow Dental School, Glasgow, UK. m.riggio@dental.gla.ac.uk
Lennon
Alan
A
D016866:Bacteroidaceae Infections / Q000175:diagnosis*; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D010482:Periapical Abscess / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D018720:Prevotella / Q000235:genetics / Q000302:isolation & purification*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013492:Suppuration / Q000382:microbiology
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S