Transactions of the Royal Society of Tropical Medicine and Hygiene (2004) 98, 148—151 Real-time polymerase chain reaction diagnosis of leishmaniasis in Panama from both fresh and frozen tissue Glenn W. Wortmann a, * , L.I. Romero b , H.M. Paz c , E. Ortega-Barria b , V. Bayard c , L.P.Hochberg d , J.R. Ryan d a Infectious Diseases Division, Ward 63, ID Clinic, Walter Reed Army Medical Center, 6900 Georgia Avenue, NW, Washington, DC 20307-5001, USA b Florida State University, Institute for Tropical Medicine and Health Science, 4750 Collegiate Drive, Panama City, Panama c Gorgas Memorial Institute for Health Studies, Panama City, Panama d Department of Entomology, Division of Communicable Diseases and Immunology, Walter Reed Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD 20910, USA Received 20 January 2003; received in revised form 5 June 2003; accepted 23 June 2003 KEYWORDS Leishmania; Diagnosis; PCR; Panama Summary Skin biopsies stored in ethanol from 49 patients with suspected cutaneous leishmaniasis (CL) were tested in a real-time polymerase chain reaction (PCR) assay and compared with conventional diagnostic methods. With clinical diagnosis as the gold standard, PCR had a sensitivity of 96% (47/49) vs. 61% (30/49) for histopathology and 33% (16/49) for culture. In addition, DNA was extracted from 70 frozen smears of lesions from suspected cases of CL and tested with the same assay. In these samples, the PCR had a sensitivity of 61% (43/70) vs. 56% (39/70) for histopathology and 41% (29/70) for culture. In this study, real-time PCR offered a rapid diagnosis with an enhanced sensitivity over conventional methods. Although the yield of PCR diagnosis waslowerwhentestingfrozensmears,theassaystilloutperformedexistingdiagnostic modalities. Published by Elsevier Ltd on behalf of Royal Society of Tropical Medicine and Hygiene. 1. Introduction Protozoans of the genus Leishmania cause cuta- neous leishmaniasis (CL), and infection with these parasites manifests into a clinical spectrum that ranges from asymptomatic infection to self-healing cutaneous lesions to severe, mutilating mucocuta- *Corresponding author. Tel.: +1-202-782-6740; fax: +1-202-782-3765. E-mail address: glenn.wortmann@na.amedd.army.mil (G.W. Wortmann). neous disease. In Panama, the first case of human cutaneous disease was reported in 1910 (Darling, 1910). The predominant parasite causing cuta- neous disease in Panama is Leishmania (Viannia) panamensis, although infection with strains of the L. mexicana complex (both L. mexicana mexicana and L. mexicana amazonensis) has been reported (de Vasquez et al., 1990; Takafugi et al., 1980). Traditional diagnosis of CL relies on biopsy of skin lesions followed by microscopic examination to identify the presence of the amastigote form of the parasite and culture to attempt propagation of 0035-9203/$ – see front matter Published by Elsevier Ltd on behalf of Royal Society of Tropical Medicine and Hygiene. doi:10.1016/S0035-9203(03)00020-8