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 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.
Teng
L J
LJ
School of Medical Technology, National Taiwan University College of Medicine, Taipei, Taiwan. ltjeng@ha.mc.ntu.edu.tw
Hsueh
P R
PR
Tsai
J C
JC
Chiang
F L
FL
Chen
C Y
CY
Ho
S W
SW
Luh
K T
KT
D001439:Bacteroides / Q000145:classification* / Q000235:genetics*; D001442:Bacteroides Infections / Q000382:microbiology*; D017931:DNA Primers; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D013045:Species Specificity
D017931:DNA Primers