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Journal of Clinical Microbiology, May 2009, p. 1386-1392, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02225-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identification of Legionella Species by Use of an Oligonucleotide Array{triangledown} ,{dagger}

Hsun-Pi Su,1,2 Seng-Kai Tung,3 Lei-Ron Tseng,2 Wen-Cherng Tsai,4 Tung-Ching Chung,2 and Tsung Chain Chang5*

Centers for Disease Control, Department of Health, Taipei, Taiwan,1 Department of Oral Hygiene, College of Health Care, China Medical University, Taichung, Taiwan,2 The Central Branch Office, Centers for Disease Control, Taichung, Taiwan,3 Super Laboratory, Taipei, Taiwan,4 Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan5

Received 19 November 2008/ Returned for modification 14 January 2009/ Accepted 25 February 2009

The genus Legionella contains a diverse group of motile, asaccharolytic, nutritionally fastidious gram-negative rods. Legionella pneumophila is the most important human pathogen, followed by L. micdadei, L. longbeachae, L. dumoffii, and other rare species. Accurate identification of Legionella spp. other than L. pneumophila is difficult because of biochemical inertness and phenotypic identity of different species. The feasibility of using an oligonucleotide array for identification of 18 species of Legionella was evaluated in this study. The method consisted of PCR amplification of the macrophage infectivity potentiator mip gene, followed by hybridization of the digoxigenin-labeled PCR products to a panel of 30 oligonucleotide probes (16- to 24-mers) immobilized on a nylon membrane. A collection of 144 target strains (strains we aimed to identify) and 50 nontarget strains (44 species) were analyzed by the array. Both test sensitivity (144/144 strains) and specificity (50/50 strains) of the array were 100%. The whole procedure for identification of Legionella species by the array can be finished within a working day, starting from isolated colonies. It was concluded that species identification of clinically relevant Legionella spp. by the array method is very reliable and can be used as an accurate alternative to conventional or other molecular methods for identification of Legionella spp.

* Corresponding author. Mailing address: Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, 1 University Rd., Tainan 701, Taiwan. Phone: 886-6-2353535, ext. 5790. Fax: 886-6-2363956. E-mail: tsungcha{at}mail.ncku.edu.tw

{triangledown} Published ahead of print on 4 March 2009.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, May 2009, p. 1386-1392, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02225-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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