Affiliation:
1. FDACenter for Food Safety and Applied Nutrition, CollegePark
2. FDA Center for BiologicsEvaluation and Research, Rockville,Maryland
Abstract
ABSTRACT
DNA
microarrays are an excellent potential tool for clinical microbiology,
since this technology allows relatively rapid identification and
characterization of microbial and viral pathogens. In the present
study, an oligonucleotide microarray was developed and used for the
analysis of thermophilic
Campylobacter
spp., the primary
food-borne pathogen in the United States. We analyzed four
Campylobacter
species:
Campylobacter jejuni
,
C.
coli
,
C. lari
, and
C. upsaliensis.
Our assay
relies on the PCR amplification of specific regions in five target
genes (
fur
,
glyA
,
cdtABC
,
ceuB-C
,
and
fliY
) as a first step, followed by microarray-based
analysis of amplified DNAs. Alleles of two genes,
fur
and
glyA
, which are found in all tested thermophilic
Campylobacter
spp., were used for identification and
discrimination among four bacterial species, the
ceuB-C
gene
was used for discrimination between
C. jejuni
and
C.
coli
, and the
fliY
and
cdt
genes were used as
additional genetic markers specific either for
C. upsaliensis
and
C. lari
or for
C. jejuni.
The array was developed
and validated by using 51 previously characterized
Campylobacter
isolates. All isolates were unambiguously
identified on the basis of hybridization patterns with 72 individual
species-specific oligoprobes. Microarray identification of
C.
jejuni
and
C. coli
was confirmed by PCR amplification of
other genes used for identification (
hipO
and
ask
).
Our results demonstrate that oligonucleotide microarrays are suitable
for rapid and accurate simultaneous differentiation among
C.
jejuni
,
C. coli
,
C. lari
, and
C.
upsaliensis
.
Publisher
American Society for Microbiology
Cited by
89 articles.
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