Novel human-associated Lachnospiraceae genetic markers improve detection of fecal pollution sources in urban waters.

Novel human-associated Lachnospiraceae genetic markers improve detection of fecal pollution sources in urban waters. Appl Environ Microbiol. 2018 May 04;: Authors: Feng S, Bootsma M, McLellan SL Abstract The human microbiome contains many organisms that could potentially be used as indicators of human fecal pollution. Here we report the development of two novel human-associated genetic marker assays that target organisms within the family Lachnospiraceae Next-generation sequencing of the V6 region of the 16S rRNA gene from sewage and animal stool samples identified 40 human-associated marker candidates with a robust signal in sewage and low or no occurrence in nonhuman hosts. Two were chosen for quantitative PCR (qPCR) assay development using longer sequences (V2 to V9 regions) generated from clone libraries. Validation of these assays, designated Lachno3 and Lachno12, was performed using fecal samples (n=55) from cat, dog, pig, cow, deer, and gull sources, and compared with established host-associated assays (Lachno2, and two Human Bacteroides assays; HB and HF183/BacR287). Each of the established assays cross-reacted with at least one other animal, including animals common in urban areas. Lachno3 and Lachno12 were primarily human-associated; however, Lachno12 demonstrated low levels of cross-reactivity with select cows, and non-specific amplification in pigs. This limitation may not be problematic when testing urban waters. These n...
Source: Applied and Environmental Microbiology - Category: Microbiology Authors: Tags: Appl Environ Microbiol Source Type: research