Kobra Mokhtarian1,2, Lame Akhlaghi1, Mohsen Mohammadi2, Ahmad Reza Meamar1, Elham Razmjou1, Majid Khoshmirsafa2,3, Reza Falak4,3. 1. Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran. 2. Immunology Research center, Iran University of Medical Sciences, Tehran, Iran. 3. Department of Immunology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran. 4. Immunology Research center, Iran University of Medical Sciences, Tehran, Iran falak.r@iums.ac.ir.
Abstract
BACKGROUND: Coprological examinations are commonly used for diagnosis of fasciolosis. However, these methods are not useful during the acute phase of the infection and also show poor sensitivity during its chronic phase. In this study we compared the immunoreactivity of the native and recombinant forms of Fasciola hepatica excretory/secretory antigens and determined the most appropriate one for development of F. hepatica-specific immunoassays. METHODS: The coding sequences of previously-determined immunogenic proteins including cathepsin L1 (CL1), fatty acid binding protein (FABP) and glutathione-S-transferase (GST) were cloned and expressed in E. coli BL-21 cells. Native forms of FABP and GST were also purified. We evaluated the immunoreactivity of the native and recombinant proteins by ELISA using sera from 40 healthy individuals, 15 fasciolosis patients, and 57 patients with other infectious diseases. RESULTS: All of the studied proteins showed high sensitivity and specificity for F. hepatica serodiagnosis. However, CL1 was more sensitive and specific (100%) than the others for the detection of F. hepatica-specific antibodies. Notably, both FABP and GST showed significant cross-reactivity with hydatidosis patients' sera while CL1 did not. CONCLUSIONS: Cathepsin L1 has acceptable sensitivity and specificity for serodiagnosis of F. hepatica and its application could be advantageous in immunoassay development.
BACKGROUND: Coprological examinations are commonly used for diagnosis of fasciolosis. However, these methods are not useful during the acute phase of the infection and also show poor sensitivity during its chronic phase. In this study we compared the immunoreactivity of the native and recombinant forms of Fasciola hepatica excretory/secretory antigens and determined the most appropriate one for development of F. hepatica-specific immunoassays. METHODS: The coding sequences of previously-determined immunogenic proteins including cathepsin L1 (CL1), fatty acid binding protein (FABP) and glutathione-S-transferase (GST) were cloned and expressed in E. coli BL-21 cells. Native forms of FABP and GST were also purified. We evaluated the immunoreactivity of the native and recombinant proteins by ELISA using sera from 40 healthy individuals, 15 fasciolosispatients, and 57 patients with other infectious diseases. RESULTS: All of the studied proteins showed high sensitivity and specificity for F. hepatica serodiagnosis. However, CL1 was more sensitive and specific (100%) than the others for the detection of F. hepatica-specific antibodies. Notably, both FABP and GST showed significant cross-reactivity with hydatidosispatients' sera while CL1 did not. CONCLUSIONS:Cathepsin L1 has acceptable sensitivity and specificity for serodiagnosis of F. hepatica and its application could be advantageous in immunoassay development.