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Journal of Clinical Microbiology, Mar 1995, 701-705, Vol 33, No. 3
H Schmidt, C Knop, S Franke, S Aleksic, J Heesemann and H Karch
In this study, we determined the sequence of the EcoRI-PstI fragment of the
plasmid pCVD432, also termed the enteroaggregative Escherichia coli
(EAggEC) probe. A primer pair complementary to this probe was designed for
PCR amplification of a 630-bp region. Comparison of the analysis of the
EAggEC probe sequence with those in database libraries revealed no
significant similarity to any known bacterial gene. Pure cultures of E.
coli cells, as well as mixed cultures from stool specimens, were
investigated with the PCR assay, the EAggEC probe test, and the adherence
test. Of 50 E. coli strains which demonstrated aggregative adherence to
HEp-2 cells, 43 (86%) were positive with the EAggEC PCR. All 43 of these
strains reacted with the EAggEC probe. Six EAggEC strains gave negative
results by both molecular techniques. In contrast, only 4 of 418 (0.96%)
strains representing other categories of diarrheagenic E. coli demonstrated
a positive PCR result. The PCR was also successful in screening for the
presence of EAggEC in enriched cultures grown from stool specimens.
Compared with cell culture assays and colony hybridization, our findings
revealed that the PCR assay was more rapid, simple, and highly sensitive
and can therefore be recommended as a screening method for EAggEC in the
clinical laboratory.
Copyright © 1995 by the American Society for Microbiology. All rights reserved.
Development of PCR for screening of enteroaggregative Escherichia coli
Institut fur Hygiene und Mikrobiologie, Universitat Wurzburg, Germany.
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