Evaluation of molecular methods for serotyping shigella flexneri

Amy Gentle, Philip M. Ashton, Tim Dallman, Claire Jenkins*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

Shigella flexneri can be phenotypically serotyped using antisera raised to type-specific somatic antigens and group factor antigens and genotypically serotyped using PCR targeting O-antigen synthesis or modification genes. The aim of this study was to evaluate a real-time PCR for serotyping S. flexneri and to use whole-genome sequencing (WGS) to investigate the phenotypic and genotypic serotype identifications. Of the 244 cultures tested retrospectively, 226 (92.6%) had concordant results between phenotypic serotyping and PCR. Seventy of the 244 isolates (including 15 of the 18 isolates where a serotype-PCR mismatch was identified) were whole-genome sequenced, and the serotype was derived from the genome. Discrepant results between the phenotypic and genotypic tests were attributed to insertions/deletions or point mutations identified in O-antigen synthesis or modification genes, rendering them dysfunctional; inconclusive serotyping results due to nonspecific cross-reactions; or novel genotypes. Phylogenetic analysis of the WGS data indicated that the serotype, regardless of whether it was phenotypically or genotypically determined, was a weak predictor of phylogenetic relationships between strains of S. flexneri. WGS data provided both genome-derived serotyping, thus supporting backward compatibility with historical data and facilitating data exchange in the community, and more robust and discriminatory typing at the single-nucleotide-polymorphism level.

Original languageEnglish
Pages (from-to)1456-1461
Number of pages6
JournalJournal of Clinical Microbiology
Volume54
Issue number6
DOIs
Publication statusPublished - Jun 2016

Bibliographical note

Funding Information:
This work, including the efforts of Amy Gentle, Philip Ashton, Timothy J. Dallman, and Claire Jenkins, was funded by DH | National Institute for Health Research (NIHR) (#109524).

Publisher Copyright:
© 2016, American Society for Microbiology. All Rights Reserved.

Fingerprint

Dive into the research topics of 'Evaluation of molecular methods for serotyping shigella flexneri'. Together they form a unique fingerprint.

Cite this