Improved in-house ELISA assay for serodiagnosis of Leishmania donovani induced cutaneous leishmaniasis in Sri Lanka

Abstract

Introduction: In Sri Lanka, cutaneous leishmaniasis (CL) is caused by a genetic variant of a visceralizing parasitic species called Leishmania donovani. A high seroprevalence of 82% has been observed in Sri Lanka during a previous study. The National Action Plan of Leishmaniasis Control of Sri Lanka and the WorldHealth Organization have recognized the significance of the development of diagnostic methods in disease control. Sensitive serodiagnostic methods will minimise the cost, need for second-line investigations, and need for methods that require invasive sampling. In this study, the diagnostic accuracy of a previously developed ELISA assay was improved further. Methods: A total of 100 samples were used (50 laboratory confirmed CL, 50 controls). Modified and previously used methods were utilised. The composition of the cell lysis buffer, blocking buffer, concentration of primary antibody and incubation periods were modified. Standard statistical methods using SPSS were used for data analysis and validation.