title: Identification of pathogenic Helicobacter species by chaperonin-60 differentiation on plastic DNA arrays.
abstract: A microarray method for bacterial species identification based on cpn60 and 16S rDNA hybridization was developed. Specific cpn60 or 16S rDNA oligonucleotides from various Helicobacter or Campylobacter species were printed and immobilized onto a proprietary plastic solid support. Using universal primers, fragments derived from either cpn60 or 16S rDNA genes from single isolates or from a complex human waste sludge DNA sample spiked with Helicobacter pylori were biotinylated and hybridized to the plastic slide. Subsequent querying with a streptavidin-horseradish peroxidase conjugate followed by color development using tetramethylbenzidine resulted in accurate Helicobacter species identification with no cross-hybridization to either the 16S rDNA or the cpn60 sequence of a closely related strain of Campylobacter jejuni. The combination of a nonfluorescence visual detection system with a polymer-based DNA microarray slide has resulted in a molecular tool that should prove useful in numerous applications requiring rapid, low-cost bacterial species identification.
authors.affiliation: Biotechnology Research Institute, National Research Council of Canada, Montréal, QC, Canada. luke.masson@nrc-cnrc.gc.ca
authors.forename: Luke
authors.initials: L
authors.lastname: Masson
authors.forename: Christine
authors.initials: C
authors.lastname: Maynard
authors.forename: Roland
authors.initials: R
authors.lastname: Brousseau
authors.forename: Swee-Han
authors.initials: SH
authors.lastname: Goh
authors.forename: Sean M
authors.initials: SM
authors.lastname: Hemmingsen
authors.forename: Janet E
authors.initials: JE
authors.lastname: Hill
authors.forename: Ana
authors.initials: A
authors.lastname: Paccagnella
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Oda
authors.forename: Naoki
authors.initials: N
authors.lastname: Kimura
mesh_terms: D016123:Campylobacter jejuni / Q000235:genetics; D018834:Chaperonin 60 / Q000032:analysis / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics*; D016998:Helicobacter / Q000235:genetics*; D006824:Hybridization, Genetic; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics*; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D018834:Chaperonin 60; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S