PCR Assay for Species-Specific Identification of Bacteroides thetaiotaomicron

Abstract

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 of B. thetaiotaomicron. Specific primers were based on shared amplicons of about 1.2 kb generated from B. 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 of B. 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 in B. thetaiotaomicron. This simple and rapid PCR assay provides a rapid and accurate method for identification of B. thetaiotaomicron and shows promise for the detection of B. thetaiotaomicron in clinical samples.

The gut microbiota instructs the hepatic endothelial cell transcriptome

“…Notably, B. thetaiotaomicron was chosen for monocolonization, as this bacterium induces angiogenesis and vascular remodeling in the small intestine ( Stappenbeck et al., 2002 ). Monocolonization of GF C57BL/6J mice was confirmed by PCR-amplification of a 721 bp product with B. thetaiotaomicron -specific 16S rDNA primers ( Figure S3 A and S3B) ( Teng et al., 2000 ). However, monocolonization of GF mice was insufficient to suppress these components of the hepatic endothelial sphingolipid metabolism ( Figure 3 C).…”