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DIAGNOSIS OF CUTANEOUS LEISHMANIASIS IN GUATEMALA USING A REAL-TIME POLYMERASE CHAIN REACTION ASSAY AND THE SMARTCYCLER^®

The polymerase chain reaction (PCR) assay has been reported for the diagnosis of cutaneous leishmaniasis (CL). Real-time (RT) PCR offers several advantages over traditional PCR, including faster processing time and decreased risk of contamination. Enhanced portability is another benefit that expands the applicability of the assay. A portable RT-PCR assay was transported from the United States to Guatemala for comparison with traditional diagnostic modalities. With the clinical diagnosis of CL as the gold standard, RT-PCR was positive in 86% (37 of 43) versus 53% (20 of 38) for microscopy and 72% (28 of 39) for culture. Negative RT-PCR samples (6) were also negative by traditional diagnostic methods (although subsequently determined to be positive by a nested kDNA PCR). Sixty-four percent (9 of 14) of cases tested and negative by microscopy and/or culture were positive by RT-PCR. This study demonstrates that a RT-PCR assay can be successfully deployed to offer enhanced sensitivity for the diagnosis of CL.

Received September 6, 2006. Accepted for publication December 5, 2006.

Acknowledgment: The authors thank Karen Collins for assisting in the DNA extraction procedures.

Financial support: The work was supported in part by funding received from the U.S. Department of Defense Gulf War Illnesses Research Program (PE 0601105D8Z).

Disclaimer: The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the U.S. Department of the Army or the U.S. Department of Defense.

^* Address correspondence to Glenn Wortmann, Ward 63, ID Clinic, Walter Reed Army Medical Center, 6900 Georgia Avenue, NW Washington, DC 20307-5001. E-mail: Glenn.Wortmann{at}na.amedd.army.mil

Authors’ addresses: Glenn Wortmann, Ward 63, Infectious Diseases Division, Walter Reed Army Medical Center, 6900 Georgia Avenue, NW Washington, DC 20307-5001, Telephone: +1 (202) 782-6740, Fax: +1 (202) 782-3765, E-mail: Glenn.Wortmann{at}na.amedd.army.mil. Lisa P. Hochberg, Department of Entomology, Division of Communicable Diseases and Immunology Walter Reed Army Institute of Research, Silver Spring, MD. Byron A. Arana, Nidia R. Rizzo, and Flora Arana, Medical Entomology Research and Training Unit, Universidad del Valle de Guatemala, Guatemala City, Guatemala. Jeffrey R. Ryan, Department of Emergency Management, Jacksonville State University, 700 Pelham Road North, Jacksonville, AL 36265.