title: Identification of Staphylococcus spp. and detection of mecA by an oligonucleotide array.
abstract: Phenotypic identification of coagulase-negative staphylococci (CoNS) is difficult and many staphylococcal species carry mecA. This study developed an array that was able to detect mecA and identify 30 staphylococcal species by targeting the internal transcribed spacer regions. A total of 129 target reference strains (30 species) and 434 clinical isolates of staphylococci were analyzed. Gene sequencing of 16S rRNA, gap or tuf genes was the reference method for species identification. All reference strains (100%) were correctly identified, while the identification rates of clinical isolates of S. aureus and CoNS were 98.9% and 98%, respectively. The sensitivity and specificity for mecA detection were 99% and 100%, respectively, in S. aureus isolates, and both values were 100% in isolates of CoNS. The assay takes 6 h from a purified culture isolate, and so far it has not been performed directly on patient samples.
authors.affiliation: Institute of Biomedical Engineering, College of Engineering, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Huan Wen
authors.initials: HW
authors.lastname: Han
authors.affiliation: Institute of Biomedical Engineering, College of Engineering, National Cheng Kung University, Tainan, Taiwan; Medical Device Innovation Center, National Cheng Kung University, Tainan, Taiwan. Electronic address: hcchang@mail.ncku.edu.tw.
authors.forename: Hsien Chang
authors.initials: HC
authors.lastname: Chang
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan. Electronic address: tsungcha@mail.ncku.edu.tw.
authors.forename: Tsung Chain
authors.initials: TC
authors.lastname: Chang
mesh_terms: D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D016106:Methicillin Resistance; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013203:Staphylococcal Infections / Q000175:diagnosis / Q000382:microbiology; D013210:Staphylococcus / Q000145:classification* / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S
keywords: Array; Coagulase-negative staphylococci; Identification; Internal transcribed spacer; Staphylococcus; mecA