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...

Full description

Saved in:
Bibliographic Details
Published in:Journal of clinical microbiology 2018-06, Vol.56 (6)
Main Authors: Banjo, Masaya, Iguchi, Atsushi, Seto, Kazuko, Kikuchi, Taisei, Harada, Tetsuya, Scheutz, Flemming, Iyoda, Sunao
Format: Article
Language:English
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
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
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 .
ISSN:0095-1137
1098-660X
DOI:10.1128/jcm.00190-18