Affiliation:
1. U.S. Department of Agriculture, Agricultural Research Service, Produce Safety and Microbiology Research Unit, Albany, California 94710
Abstract
ABSTRACT
To explore the use of DNA microarrays for pathogen detection in food, we produced DNA oligonucleotide arrays to simultaneously determine the presence of
Arcobacter
and the presence of
Campylobacter
in retail chicken samples. Probes were selected that target housekeeping and virulence-associated genes in both
Arcobacter butzleri
and thermotolerant
Campylobacter jejuni
and
Campylobacter coli
. These microarrays showed a high level of probe specificity; the signal intensities detected for
A. butzleri
,
C. coli
, or
C. jejuni
probes were at least 10-fold higher than the background levels. Specific identification of
A. butzleri
,
C. coli
, and
C. jejuni
was achieved without the need for a PCR amplification step. By adapting an isolation method that employed membrane filtration and selective media,
C. jejuni
isolates were recovered from package liquid from whole chicken carcasses prior to enrichment. Increasing the time of enrichment resulted in the isolation of
A. butzleri
and increased the recovery of
C. jejuni. C. jejuni
isolates were further classified by using an additional subset of probes targeting the lipooligosaccharide (LOS) biosynthesis locus. Our results demonstrated that most of the
C. jejuni
isolates likely possess class B, C, or H LOS. Validation experiments demonstrated that the DNA microarray had a detection sensitivity threshold of approximately 10,000
C. jejuni
cells. Interestingly, the use of
C. jejuni
sequence-specific primers to label genomic DNA improved the sensitivity of this DNA microarray for detection of
C. jejuni
in whole chicken carcass samples.
C. jejuni
was efficiently detected directly both in package liquid from whole chicken carcasses and in enrichment broths.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
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