Analytical comparison of nine PCR primer sets designed to detect the presence of Escherichia coli/Shigella in water samples.
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.
Maheux
Andrée F
AF
Centre de recherche en infectiologie de l'Université Laval, Centre hospitalier universitaire de Québec (Pavillon CHUL), 2705 Laurier Blvd., Suite RC-709, Québec (Québec), Canada G1V 4G2.
Picard
François J
FJ
Boissinot
Maurice
M
Bissonnette
Luc
L
Paradis
Sonia
S
Bergeron
Michel G
MG
D017931:DNA Primers / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012760:Shigella / Q000235:genetics / Q000302:isolation & purification*; D014871:Water Microbiology
D017931:DNA Primers