4.8 Article

Analytical comparison of nine PCR primer sets designed to detect the presence of Escherichia coli/Shigella in water samples

Journal

WATER RESEARCH
Volume 43, Issue 12, Pages 3019-3028

Publisher

PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.watres.2009.04.017

Keywords

Escherichia coli; PCR; Primers; Shigella; Target gene

Funding

  1. Canadian Institutes of Health Research (CIHR) [PA-15586]
  2. Canadian Foundation for Innovation (CFI) [FCI-5251]
  3. Nasivvik (Center for Inuit Health and Changing Environment
  4. Canadian Institutes for Health Research)

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The analytical performance of 9 different PCR primer sets designed to detect Escherichia coli and Shigella in water has been evaluated in terms of ubiquity, specificity, and analytical detection limit. Of the 9 PCR primer sets tested, only 3 of the 5 primer sets targeting uidA gene and the primer set targeting tuf gene amplified DNA from all E. coli strains tested. However, of those 4 primer sets, only the primer set targeting the tuf gene also amplified DNA from all Shigella strains tested. For the specificity, only the primer sets targeting the uidA gene were 100% specific although the primer sets targeting 16S rRNA, phoE, and tuf genes only amplified Escherichia fergusonii as non-specific target. Finally, the primer set targeting the 16S-ITS-23S gene region, was not specific as it amplified DNA from many other Enterobacteriaceae species. In summary, only the assay targeting the tuf gene detected all E. coli/Shigella strains tested in this study. However, if it becomes important to discriminate between E. coli and E. fergusonii, assays targeting the uidA gene would represent a good choice although none of them were totally ubiquitous to detect of the presence of Shigella strains. (C) 2009 Elsevier Ltd. All rights reserved.

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