Evaluation of genetic markers from the 16S rRNA gene V2 region for use in quantitative detection of selected Bacteroidales species and human fecal waste by qPCR

Systematic and Applied Microbiology
Richard A HauglandOrin C Shanks

Abstract

Molecular methods for quantifying defined Bacteroidales species from the human gastrointestinal tract may have important clinical and environmental applications, ranging from diagnosis of infections to fecal source tracking in surface waters. In this study, sequences from the V2 region of the small subunit ribosomal RNA gene were targeted in the development of qPCR assays to quantify DNA from six Bacteroides and one Prevotella species. In silico and experimental analyses suggested that each of the assays was highly discriminatory in detecting DNA from the intended species. Analytical sensitivity, precision and ranges of quantification were demonstrated for each assay by coefficients of variation of less than 2% for cycle threshold measurements over a range from 10 to 4×10(4) target sequence copies. The assays were applied to assess the occurrence and relative abundance of their target sequences in feces from humans and five animal groups as well as in 14 sewage samples from 13 different treatment facilities. Sequences from each of the species were detected at high levels (>10(3)copies/ng total extracted DNA) in human wastes. Sequences were also detected by each assay in all sewage samples and, with exception of the Prevotella s...Continue Reading

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Jul 14, 2009·Applied and Environmental Microbiology·Orin C ShanksRichard A Haugland

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Citations

Sep 10, 2014·Trends in Microbiology·Sandra L McLellan, A Murat Eren
Jun 7, 2011·Applied and Environmental Microbiology·Shane W RogersOrin C Shanks
Apr 17, 2012·Applied and Environmental Microbiology·Catherine A KeltyOrin C Shanks
Nov 12, 2013·Applied and Environmental Microbiology·Kruti RavaliyaLee-Ann Jaykus
Jul 25, 2013·Water Research·Alexandria B BoehmStephen B Weisberg
Apr 2, 2013·Canadian Journal of Microbiology·Corinne A KrentzJudith Isaac-Renton
Feb 5, 2016·Journal of Microbiological Methods·Richard A HauglandChristopher Sinigalliano
Sep 3, 2011·Environmental Microbiology·Hyatt C GreenKatharine G Field
Apr 6, 2016·Environmental Science & Technology·Angela HarrisAlexandria B Boehm
Nov 2, 2014·Environmental Microbiology Reports·Amber M KoskeySandra L McLellan
Jul 21, 2015·Applied Microbiology and Biotechnology·Ann M ArfkenMichael A Mallin
Dec 18, 2013·The Science of the Total Environment·Chang Soo LeeJiyoung Lee
Dec 22, 2016·Environmental Science & Technology·Patrick R RoehrdanzPatricia A Holden
Jun 20, 2017·Journal of Microbiological Methods·Laura SeidelAndreas Nocker
Jul 13, 2017·Environmental Science & Technology·Elyse StachlerOrin C Shanks
Sep 20, 2017·Microbial Biotechnology·Cristina García-AljaroJuan Jofre
Feb 7, 2018·Environmental Science. Processes & Impacts·Wiley C JenningsAlexandria B Boehm
Dec 1, 2017·Journal of Molecular Microbiology and Biotechnology·Liliana Pampillón-GonzálezYendi E Navarro-Noya
May 8, 2018·Applied and Environmental Microbiology·Shuchen FengSandra L McLellan
Jun 5, 2018·Microbes and Environments·Hye Young KoGwangPyo Ko
Sep 17, 2013·Angewandte Chemie·Yulia V Gerasimova, Dmitry M Kolpashchikov
Jun 10, 2016·Journal of Water and Health·Jessica Healy-ProfitósJiyoung Lee

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