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Use of a Bacteroides thetaiotaomicron ‐specific α‐1‐6, mannanase quantitative PCR to detect human faecal pollution in water
Author(s) -
YamparaIquise H.,
Zheng G.,
Jones J.E.,
Carson C. Andrew
Publication year - 2008
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2008.03895.x
Subject(s) - bacteroides thetaiotaomicron , biology , bacteroides , sewage , feces , bacteroidaceae , microbiology and biotechnology , bacteria , gene , genetics , environmental science , environmental engineering
Aims: 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. Methods and Results: qPCR primers, based on the complete genomic sequence of B. thetaiotaomicron VPI 5482, were designed and tested. The single‐copy putative mannanase homologue, α‐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 × 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. Conclusion: The subject assay is potentially useful for quantification of sewage pollution in water. Significance and Impact of the Study: 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.