PCR Assay for Species-Specific Identification of Bacteroides thetaiotaomicron

Bacteroides thetaiotaomicron is the second most frequently encountered species of the anaerobes isolated from clinical specimens. We developed a PCR-based assay for the rapid identification ofB. thetaiotaomicron . Specific primers were based on shared amplicons of about 1.2 kb generated fromB. thetaiotaomicron by randomly amplified polymorphic DNA. This 1.2-kb fragment was sequenced and then used to design a set of PCR amplification primers. This PCR generated an amplification product of 721 bp, which was unique to all 65 isolates ofB. thetaiotaomicron tested. There was no amplification with isolates of other bacterial species. Restriction enzyme digestion of the amplification product and dot blot hybridization further verified the specificity of the assay. These results suggest that this PCR assay targets a nucleotide sequence that is strongly conserved inB. thetaiotaomicron . This simple and rapid PCR assay provides a rapid and accurate method for identification ofB. thetaiotaomicron and shows promise for the detection ofB. thetaiotaomicron in clinical samples.