Utilizing novel Escherichia coli ‐specific conserved signature proteins for enhanced monitoring of recreational water quality

This paper explores the application of a novel DNA sequence type, specifically conserved signature genes encoding conserved signature proteins (CSPs), for bacterial identification. Although our focus is on the identification ofEscherichia coli, the same principle can be extended to identify other taxa using their respective CSPs. AbstractEscherichia coli serves as a proxy indicator of fecal contamination in aquatic ecosystems. However, its identification using traditional culturing methods can take up to 24  h. The application of DNA markers, such as conserved signature proteins (CSPs) genes (unique to all species/strains of a specific taxon), can form the foundation for novel polymerase chain reaction (PCR) tests that unambiguously identify and detect targeted bacterial taxa of interest. This paper r eports the identification of three new highly-conserved CSPs (genes), namelyYahL,YdjO, andYjfZ, which are exclusive toE. coli/Shigella. Using PCR primers based on highly conserved regions within these CSPs, we have developed  quantitative PCR (qPCR) assays for the evaluation ofE. coli/Shigella species in water ecosystems. Both in-silico and experimental PCR testing confirmed the absence of sequence match when tested against other bacteria, thereby confirming 100% specificity of the tested CSPs forE. coli/Shigella. The qPCR assays for each of the three CSPs provided reliable quantification for all tested enterohaemorrhagic and environmentalE. coli strains, a requirement for wat...
Source: MicrobiologyOpen - Category: Microbiology Authors: Tags: ORIGINAL ARTICLE Source Type: research