Journal
JOURNAL OF CLINICAL MICROBIOLOGY
Volume 56, Issue 6, Pages -Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JCM.00190-18
Keywords
Escherichia coli; PCR; flagellin; serotyping
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Funding
- Research Program on Emerging and Reemerging Infectious Diseases from the Japan Agency for Medical Research and Development (AMED) [JP15fk0108008]
- Grants-in-Aid for Scientific Research [15K08486] Funding Source: KAKEN
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In Escherichia coli, more than 180 O groups and 53 H types have been recognized. The O:H serotypig of E. coli strains is an effective method for identifying strains with pathogenic potential and classifying them into clonal groups. In particular, the serotyping of Shiga toxin-producing E. coli (STEC) strains provides valuable information to evaluate the routes, sources, and prevalence of agents in outbreak investigations and surveillance. Here, we present a complete and practical PCR-based H-typing system, E. coli H-genotyping PCR, consisting of 10 multiplex PCR kits with 51 single PCR primer pairs. Primers were designed based on a detailed comparative analysis of sequences from all H-antigen (flagellin)-encoding genes, fliC and its homologs. The specificity of this system was confirmed by using all H type reference strains. Additionally, 362 serotyped wild strains were also used to evaluate its practicality. All 277 H-type-identified isolates gave PCR products that corresponded to the results of serological H typing. Moreover, 76 nonmotile and nine untypeable strains could be successfully subtyped into any H type by the PCR system. The E. coli H-genotyping PCR developed here allows broader, rapid, and low-cost subtyping of H types and will assist epidemiological studies as well as surveillance of pathogenic E. coli.
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