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Journal of Clinical Microbiology, October 2006, p. 3510-3516, Vol. 44, No. 10
0095-1137/06/$08.00+0     doi:10.1128/JCM.00670-06

Species-Specific Identification of Leptospiraceae by 16S rRNA Gene Sequencing

Roger E. Morey, Renee L. Galloway, Sandra L. Bragg, Arnold G. Steigerwalt, Leonard W. Mayer, and Paul N. Levett*

Meningitis and Special Pathogens Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia

Received 29 March 2006/ Returned for modification 10 April 2006/ Accepted 20 July 2006

The genus Leptospira is classified into 13 named species and 4 genomospecies based upon DNA-DNA reassociation studies. Phenotypic tests are unable to distinguish between species of Leptospira, and there is a need for a simplified molecular approach to the identification of leptospires. 16S rRNA gene sequences are potentially useful for species identification of Leptospira, but there are a large number of sequences of various lengths and quality in the public databases. 16S rRNA gene sequences of near full length and bidirectional high redundancy were determined for all type strains of the species of the Leptospiraceae. Three clades were identified within the genus Leptospira, composed of pathogenic species, nonpathogenic species, and another clade of undetermined pathogenicity with intermediate 16S rRNA gene sequence relatedness. All type strains could be identified by 16S rRNA gene sequences, but within both pathogenic and nonpathogenic clades as few as two or three base pairs separated some species. Sequences within the nonpathogenic clade were more similar, and in most cases ≤10 bp distinguished these species. These sequences provide a reference standard for identification of Leptospira species and confirm previously established relationships within the genus. 16S rRNA gene sequencing is a powerful method for identification in the clinical laboratory and offers a simplified approach to the identification of Leptospira species.

* Corresponding author. Mailing address: Saskatchewan Health, Provincial Laboratory, 3211 Albert Street, Regina, Saskatchewan S4S 5W6, Canada. Phone: (306) 787-3135. Fax: (306) 787-1525. E-mail: plevett{at}health.gov.sk.ca.

Journal of Clinical Microbiology, October 2006, p. 3510-3516, Vol. 44, No. 10
0095-1137/06/$08.00+0     doi:10.1128/JCM.00670-06




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