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Journal of Clinical Microbiology, December 1998, p. 3492-3496, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

16S Ribosomal DNA Typing for Identification of Pathogens in Patients with Bacterial Keratitis

C. Michele Knox,1,2 Vickey Cevellos,1 and Deborah Dean1,3,*

Francis I. Proctor Foundation1 and Departments of Ophthalmology2 and Medicine, Division of Infectious Disease,3 University of California at San Francisco School of Medicine, San Francisco, California

Received 5 June 1998/Returned for modification 3 August 1998/Accepted 1 September 1998

The identification of pathogens in patients with bacterial keratitis remains problematic because standard diagnostic tests are negative for 40 to 60% of patients. A cross-sectional study was undertaken to determine if PCR and sequence analysis of 16S ribosomal DNA (rDNA) could be used to detect bacterial pathogens in patients with keratitis. Corneal specimens were collected for culture and rDNA typing. Variable segments of each rDNA specimen were amplified by PCR, sequenced, and aligned with the sequences in GenBank. Eleven patients had microbiologically documented bacterial keratitis, while 17 patients had keratitis due to other causes. Nine (82%) of 11 bacterial keratitis patients were PCR positive; each sequencing result matched the culture results. Seventeen (100%) patients with nonbacterial keratitis were PCR negative. Our data suggest that 16S rDNA typing holds promise as a rapid alternative to culture for identifying pathogens in patients with bacterial keratitis.

* Corresponding author. Mailing address: University of California at San Francisco, Box 0412, S-307, 513 Parnassus Ave., San Francisco, CA 94143-0412. Phone: (415) 476-4548. Fax: (415) 476-6085. E-mail: debd{at}itsa.ucsf.edu.

Journal of Clinical Microbiology, December 1998, p. 3492-3496, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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