Development of a duplex real-time PCR for differentiation between E. coli and Shigella spp

Aims: The aim of this study was to develop a real-time PCR test for differentiation between Shigella spp. and E. coli, in particular enteroinvasive Escherichia coli (EIEC). Methods and Results: A duplex real-time PCR specific for the genes encoding for β-glucuronidase (uidA) and lactose permease (la...

Full description

Saved in:
Bibliographic Details
Published in:Journal of applied microbiology 2011-05, Vol.110 (5), p.1245-1251
Main Authors: Pavlovic, M, Luze, A, Konrad, R, Berger, A, Sing, A, Busch, U, Huber, I
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
beta-glucuronidase
Biological and medical sciences
DNA, Bacterial - genetics
E. coli
enteroinvasive Escherichia coli
Escherichia coli - classification
Escherichia coli - genetics
Escherichia coli - isolation & purification
Fundamental and applied biological sciences. Psychology
genes
Genes, Bacterial
Glucuronidase - genetics
lactose
lacY
Limit of Detection
Membrane Transport Proteins - genetics
Microbiology
multiplex PCR
Polymerase Chain Reaction - methods
quantitative polymerase chain reaction
real‐time PCR
serotypes
Shigella
Shigella - classification
Shigella - genetics
Shigella - isolation & purification
Species Specificity
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Aims: The aim of this study was to develop a real-time PCR test for differentiation between Shigella spp. and E. coli, in particular enteroinvasive Escherichia coli (EIEC). Methods and Results: A duplex real-time PCR specific for the genes encoding for β-glucuronidase (uidA) and lactose permease (lacY) was developed. Ninety-six isolates including 11 EIEC isolates of different serotypes and at least three representatives of each Shigella species were used for selectivity testing. All isolates tested were positive for the uidA gene. Additionally, all E. coli isolates were positive for the lacY gene, whereas no Shigella isolate tested harboured lacY. Conclusions: The duplex real-time PCR assay was found to be simple, rapid, reliable and specific. Significance and Impact of the Study: If possible at all, delineation of so-called inactive EIEC from Shigella spp. is cumbersome. Biochemical and serological methods are limited to specific pheno- and serotypes. This assay clearly simplifies the differentiation of both.
ISSN:1364-5072
1365-2672
DOI:10.1111/j.1365-2672.2011.04973.x