Escherichia coli H-Genotyping PCR: a Complete and Practical Platform for Molecular H Typing

• Published in: Journal of clinical microbiology Vol. 56; no. 6
• Main Authors: Banjo, Masaya, Iguchi, Atsushi, Seto, Kazuko, Kikuchi, Taisei, Harada, Tetsuya, Scheutz, Flemming, Iyoda, Sunao
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01-06-2018
• Subjects: Antigens, Bacterial - genetics; Bacteriology; DNA Primers; DNA, Bacterial - genetics; Escherichia coli - classification; Escherichia coli - isolation & purification; Escherichia coli Infections - diagnosis; Escherichia coli Proteins - genetics; Flagellin - genetics; Genotype; Genotyping Techniques; Humans; Molecular Typing - economics; Molecular Typing - methods; Multiplex Polymerase Chain Reaction - methods; Serogroup; serotyping; flagellin; Escherichia coli; PCR
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/jcm.00190-18

In
, more than 180 O groups and 53 H types have been recognized. The O:H serotyping of
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
(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,
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,
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
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
.