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Journal of Clinical Microbiology, May 2009, p. 1458-1462, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02365-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Molecular Identification of T4 and T5 Genotypes in Isolates from Acanthamoeba Keratitis Patients{triangledown}

D. R. Ledee,1* A. Iovieno,1 D. Miller,1 N. Mandal,3 M. Diaz,2 J. Fell,2 M. E. Fini,4 and E. C. Alfonso1

Department of Ophthalmology, Bascom Palmer Eye Institute,1 Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, Florida,2 Academic Unit of Ophthalmology, University of Birmingham, Edgbaston, United Kingdom,3 Institute for Genetic Medicine, Keck School of Medicine, University of Southern California, Los Angeles, California4

Received 9 December 2008/ Returned for modification 25 February 2009/ Accepted 16 March 2009

Acanthamoeba keratitis (AK) is a rare but sight-threatening ocular infection. Outbreaks have been associated with contaminated water and contact lens wear. The epidemiology and pathology may be associated with unique genotypes. We determined the Rns genotype for 37 clinical isolates from 23 patients presenting at the University of Miami Bascom Palmer Eye Institute with confirmed AK infections in 2006 to 2008. The genus-specific ASA.S1 amplicon allowed for rapid genotyping of the nonaxenic cultures. Of the 37 isolates, 36 were of the T4 genotype. Within this group, 13 unique diagnostic fragment 3 sequences were identified, 3 of which were not in GenBank. The 37th isolate was a T5, the first in the United States and second worldwide to be found in AK. For five patients with isolates from the cornea and contact lens/case, identical sequences within each patient cluster were observed, confirming the link between contact lens contamination and AK infection. Genotyping is an important tool in the epidemiological study of AK. In this study, it allowed for the detection of new strains and provided an etiological link between source and infection. Additionally, it can allow for accurate categorizing of physiological differences, such as strain virulence, between isolates and clades.

* Corresponding author. Mailing address: Bascom Palmer Eye Institute, Miller School of Medicine, University of Miami, 1638 NW 10th Ave., Miami, FL 33136. Phone: (425) 402-1808. Fax: (801) 407-2379. E-mail: dolenal{at}yahoo.com

{triangledown} Published ahead of print on 25 March 2009.

Journal of Clinical Microbiology, May 2009, p. 1458-1462, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02365-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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