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Journal of Clinical Microbiology, December 1999, p. 3906-3911, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of Porphyromonas gingivalis Strains by Heteroduplex Analysis and Detection of Multiple Strains

Eugene J. Leys,1,* James H. Smith,2,dagger Sharon R. Lyons,1 and Ann L. Griffen3

Departments of Oral Biology1 and Pediatric Dentistry,3 College of Dentistry, and Department of Internal Medicine, College of Medicine,2 The Ohio State University, Columbus, Ohio

Received 1 February 1999/Returned for modification 27 April 1999/Accepted 4 August 1999

Heteroduplex analysis has been used extensively to identify allelic variation among mammalian genes. It provides a rapid and reliable method for determining and cataloging minor differences between two closely related DNA sequences. We have adapted this technique to distinguish among strains or clonal types of Porphyromonas gingivalis. The ribosomal intergenic spacer region (ISR) was amplified directly from a subgingival plaque sample by PCR with species-specific primers, avoiding the need for culturing the bacteria. The PCR products were then directly compared by heteroduplex analysis with known strains of P. gingivalis for identification. We identified 22 distinct but closely related heteroduplex types of P. gingivalis in 1,183 clinical samples. Multiple strains were found in 34% of the samples in which P. gingivalis was detected. Heteroduplex types were identified from these multistrain samples without separating them by culturing or molecular cloning. PCR with species-specific primers and heteroduplex analysis makes it possible to reliably and sensitively detect and identify strains of P. gingivalis in large numbers of samples.

* Corresponding author. Mailing address: Department of Oral Biology, College of Dentistry, The Ohio State University, 305 W. 12th Ave., Columbus, OH 43210. Phone: (614) 292-5273. Fax: (614) 292-6087. E-mail: leys.1{at}osu.edu.

dagger Present address: Infectious Disease Consultants, Inc., Columbus, Ohio.

Journal of Clinical Microbiology, December 1999, p. 3906-3911, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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