Use of a Bacteroides thetaiotaomicron-specific alpha-1-6, mannanase quantitative PCR to detect human faecal pollution in water.
OBJECTIVE
The aims of this work were to develop a quantitative test, based on Bacteroides thetaiotaomicron, for human faecal pollution in water and to evaluate test performance.


RESULTS
qPCR primers, based on the complete genomic sequence of B. thetaiotaomicron VPI 5482, were designed and tested. The single-copy putative mannanase homologue, alpha-1-6 mannanase, was selected as the particular target and sequences within this gene chosen as the qPCR primers by Blast search for specificity to B. thetaiotaomicron. The average concentration of B. thetaiotaomicron in human faeces was 1.39 x 10(8) cells per gram faeces and the detection limit was 9.3 B. thetaiotaomicron copies per qPCR procedure. Comparison of B. thetaiotaomicron content in sewage vs pooled nonhuman faecal samples indicated that the current assay is specific for sewage.


CONCLUSIONS
The subject assay is potentially useful for quantification of sewage pollution in water.


CONCLUSIONS
Bacteroides-associated markers, proposed for faecal source tracking, have exclusively been based on gene sequences related to generally classified and uncultured bacteria. However, genes associated with host-microbe interaction have been suggested as more specific markers. The present assay targets such a gene of B. thetaiotaomicron which is considered to be a symbiont in the human gut.
Yampara-Iquise
H
H
Department of Veterinary Pathobiology, University of Missouri, Columbia, MO, USA.
Zheng
G
G
Jones
J E
JE
Carson
C Andrew
CA
D000818:Animals; D001439:Bacteroides / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012722:Sewage / Q000382:microbiology; D014871:Water Microbiology; D014876:Water Pollution
D017931:DNA Primers; D004269:DNA, Bacterial; D012722:Sewage