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Utility of a Protein Fraction with Cathepsin L-Like Activity Purified from Cysticercus Fluid of Taenia solium in the Diagnosis of Human Cysticercosis

Neurocysticercosis, an endemic parasitic disease in most developing countries, is caused by Taenia solium and compromises the human central nervous system. Cathepsin L-like proteases are secreted by several parasites including T. solium and constitute important antigens for immunodiagnostics. A protein fraction with cathepsin L-like activity was purified from the cysticercus fluid by size exclusion and ion exchange chromatography. Cathepsin L-like activity was measured fluorometrically by detecting the hydrolysis of the fluorogenic substrate Z-Phe-Arg-AMC. The purified protein fraction included antigens of 53 and 25 kD that were tested in a Western immunoblot and in an enzyme-linked immunosorbent assay (ELISA) for detection of human cysticercosis. The sensitivity of the Western immunoblot was 96% for patients infected with multiple cysts and 78% for patients with a single cyst. Specificity was 98%. The sensitivity of the ELISA was 98% in patients with multiple cysts and 84% in patients with a single cyst. Specificity was 92.7%.

Received October 2, 2008. Accepted for publication March 6, 2009.

Acknowledgments: We thank Dr. Clinton White, Dr. Philip LoVerde, Dr. Manuela Verástegui, and Dr. Jeanette Velasquez for their advice and review of the manuscript. We are also grateful to Myra Flores and Patricia Arias for their hard work on sample processing.

Financial support: This study was partially supported by research grants P01 AI51976 TMRC from the National Institutes of Health; 23981 from The Bill and Melinda Gates Foundation; U01 AI35894, TW05562, K24 A10668903, R01 TW005860, and D43 TW007120 from the National Institutes of Health; 01107 from the Food and Drug Administration; and 063109 from The Wellcome Trust, United Kingdom.

Disclaimer: The sponsors had no role in the design or writing of this manuscript.

^* Address correspondence to Robert H. Gilman, Department of International Health, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Room W5515, Baltimore, MD 21205. E-mail: rgilman{at}jhsph.edu

Authors’ addresses: Mirko Zimic, Mónica Pajuelo, Daniel Rueda, César López, Yanina Arana, Yesenia Castillo, Maritza Calderón, and Patricia Sheen, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias, Universidad Peruana Cayeatno Heredia. Av. Honorio Delgado 430, SMP Lima 31, Perú. Silvia Rodriguez and Héctor H. García, Unidad de Cisticercosis, Instituto Nacional de Ciencias Neurológicas, Jr. Ancash 1271, Barrios Altos, Lima 1, Perú. Joseph M. Vinetz, Division of Infectious Diseases, Department of Medicine, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093. Armando Gonzales, Escuela de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Av. Circunvalación Cdra, 28 s/n - San Borja, Lima, Perú. Robert H. Gilman, Department of International Health, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Room W5515, Baltimore, MD 21205, E-mail: rgilman{at}jhsph.edu.