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

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 inf...

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Bibliographic Details
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
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Summary:	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 .
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Citation Banjo M, Iguchi A, Seto K, Kikuchi T, Harada T, Scheutz F, Iyoda S, Pathogenic E. coli Working Group in Japan. 2018. Escherichia coli H-genotyping PCR: a complete and practical platform for molecular H typing. J Clin Microbiol 56:e00190-18. https://doi.org/10.1128/JCM.00190-18.
ISSN:	0095-1137 1098-660X
DOI:	10.1128/jcm.00190-18