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J. Clin. Microbiol. doi:10.1128/JCM.01712-06
Copyright (c) 2006, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Array-based Identification of Species of Abiotrophia, Enterococcus, Granulicatella and Streptococcus

Department of Medical Laboratory Science and Biotechnology, School of Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China; School of Medical Technology, National Taiwan University College of Medicine, Taipei, Taiwan, Republic of China; Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium; Division of Clinical Microbiology, Department of Pathology, National Cheng Kung University Hospital, Tainan, Taiwan, Republic of China

^* To whom correspondence should be addressed. Email: tsungcha{at}mail.ncku.edu.tw.

	Abstract

Some species of enterococci and streptococci are difficult to differentiate by phenotypic traits. The feasibility of using an oligonucleotide array for identification of 11 viridans group streptococci was previously established. The aim of this study was to expand the array to identify species of Abiotrophia (1 species), Enterococcus (18 species), Granulicatella (3 species), and Streptococcus (31 species and 6 subspecies). The method consisted of PCR amplification of the ribosomal DNA intergenic spacer (ITS) regions, followed by hybridization of the digoxigenin-labeled PCR products to a panel of oligonucleotide probes (16- to 30-mers) immobilized on nylon membrane. Probes were could be divided into three categories: species-specific, group-specific, and supplemental probes. All probes were designed either from the ITS regions or from the 3' end of the 16S rRNA genes. A collection of 312 target (162 reference strains and 150 clinical isolates) and 73 non-target strains was identified by the array. Most clinical isolates were isolated from blood cultures or deep abscesses and only those strains having excellent species identification with the rapid ID 32 STREP system (bioMé rieux Vitek, Taipei, Taiwan) were used for array testing. The test sensitivity and specificity of the array were 100% (312/312) and 98.6% (72/73), respectively. The whole procedures of array hybridization took about 8 h starting from isolated colonies and the hybridization patterns could be read by the naked eye. The oligonucleotide array is accurate for identification of the above microorganisms and could be used as a reliable alternative of phenotypic identification methods.

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