Diagn Microbiol Infect Dis
Division of Microbiology, Osaka Institute of Public Health, Osaka, Japan.
Published: March 2023
To develop subtyping methods for Shiga toxin (Stx)1a, Stx1c, Stx1d, Stx2a, Stx2b, Stx2c, Stx2d, Stx2e, Stx2f, and Stx2g genes for epidemiological analyses of Shiga toxin-producing Escherichia coli (STEC), we developed 10 simplex real-time polymerase chain reaction (PCR) assays with reference to 284 valid stx sequences and evaluated their specificity and quantitative accuracy using STEC and non-STEC isolates and recombinant plasmids, respectively. Three stx and 5 stx subtype genes, except for stx and stx, were detected with high specificity using STEC isolates. However, some stx sequences potentially being close to both Stx2a and Stx2d cluster in neighbor-joining cluster analysis were positive for stx and stx by real-time PCR. For the stx assay, the number of real-time PCR cycles was reduced to avoid unnecessary false-positive results. Based on these considerations, the real-time PCR assays developed here might aid epidemiological investigations of infections or outbreaks caused by STEC harboring any of the stx subtype genes.
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http://dx.doi.org/10.1016/j.diagmicrobio.2022.115874 | DOI Listing |
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