title: An electronic DNA microarray technique for detection and differentiation of viable Campylobacter species.
abstract: An electronic oligonucleotide microarray technique was developed for detection and differentiation of the viable Campylobacter species, C. jejuni, C. coli, and C. lari. This development consisted of four major components: identification of single nucleotide polymorphisms (SNPs) within the hsp60 gene as species markers, design of fluorescently labelled SNP-based reporters, development of an electronic microarray detection, and application of the integrated technique to analysis of Campylobacter species in food samples. A unique capability of this technique is the specific detection of viable cells and not dead ones. This is achieved by using mRNA of the 60 kDa heat-shock protein as the viability marker. The identification of two unique SNPs closely located at positions 291 and 294 of the hsp60 gene enabled the differentiation of the three Campylobacter species. This technique was able to detect as few as two viable Campylobacter cells. The analysis of 19 blind Campylobacter samples showed 100% agreement with their identities obtained using pulsed-field gel electrophoresis. The analysis of six chicken samples revealed the presence of C. coli in one of the samples.
authors.affiliation: Environmental Health Sciences, Department of Public Health Sciences, University of Alberta, Edmonton, Alberta, CanadaT6G 2G3.
authors.forename: Hai
authors.initials: H
authors.lastname: Zhang
authors.forename: Zhilong
authors.initials: Z
authors.lastname: Gong
authors.forename: Odell
authors.initials: O
authors.lastname: Pui
authors.forename: Yanming
authors.initials: Y
authors.lastname: Liu
authors.forename: Xing-Fang
authors.initials: XF
authors.lastname: Li
mesh_terms: D000818:Animals; D002167:Campylobacter / Q000235:genetics* / Q000302:isolation & purification; D018834:Chaperonin 60 / Q000235:genetics*; D002645:Chickens; D005506:Food Contamination / Q000032:analysis*; D020869:Gene Expression Profiling / Q000379:methods*; D005798:Genes, Bacterial; D008460:Meat / Q000382:microbiology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D013045:Species Specificity
chemical_list: D018834:Chaperonin 60