Project 1-115 Indonesia

PARTNERSHIPS FOR ENHANCED ENGAGEMENT IN RESEARCH (PEER) HEALTH
Cycle 1

Development of an antigen-capture immunoassay for the rapid diagnosis of acute leptospirosis.

Principal Investigator: Farida Handayani, Ministry of Health, Indonesia

NIH-Supported Collaborator: David AuCoin, University of Nevada, Reno

Title of NIH Award: Antigen Detection Assay for the Diagnosis of Melioidosis
Project Dates: February 2016 - January 2020

Project Overview

Leptospirosis is a globally important zoonotic disease that is endemic in South East Asia with human infection commonly reported throughout the region. Leptospirosis is classified as a neglected disease in Indonesia and has been an increasing public health issue in the country. A number of outbreaks of leptospirosis have occurred in several areas in Indonesia during 2004-2012 with mortality rates as high as 35%. Laboratory diagnosis of leptospirosis is difficult due to limitations in laboratory diagnostics. Clinical diagnosis is also difficult since leptospirosis symptoms are variable, ranging from a classical flu syndrome to Weil’s disease, which typically causes major hepatic and renal failures, often leading to death. Specific laboratory tests, including bacteria isolation, DNA testing and serology using the microagglutination test (MAT), still remain limited to highly specialized laboratories. The goal of this study is to develop a sensitive, noninvasive and inexpensive immunoassay for point-of-care diagnosis of leptospirosis. The project will significantly improve the diagnosis of leptospirosis, as well as provide a better understanding of leptospirosis endemicity within Indonesia. This study will also aid and enhance the Ministry of Health’s efforts to control leptospirosis. This project has three components: 1.) Production of a library of monoclonal antibodies (mAbs) specific to LPS or other circulating antigens that are secreted or shed during infection of Leptospira spp. for the development of a prototype antigen-capture ELISA. 2.) Construction of prototype immunoassay for leptospirosis diagnosis. 3.) Conduct a prevalence survey of Leptospirosis in Endemic areas in Indonesia using the immunoassay developed in Phase 2.

Summary of Recent Activities

2017 was a year of significant progress and effort in the laboratory to detect Leptospira antigen in the urine patient samples. The team has worked hard to accurately characterize and determine whether urine samples of leptospirosis patients are truly infected by Leptospira before sending the samples to Dr. Aucoin at University of Nevada Reno. They were able to succeed by developing a protocol that allowed them to detect the very low DNA Leptospira in the urine samples and now have proven laboratory SOPs to detect Leptospira in low level concentrations, both in urine and blood or even other resources samples. This has been a major breakthrough for the team as it is very tricky to detect low level concentration of Leptospira antigen, especially in urine samples.

The team is now in the process of preparing a publication entitled: Screening Urine samples using qPCR methods by SecY gene and LipL32. Dr. Ahmed from FAO/OIE and National Collaborating Centre for Reference and Research on Leptospirosis. KIT Biomedical Research in the Netherlands has agreed to review the publication draft and the team met with them during past ILS meeting in New Zealand. In addition, The team published a book, entitled 'Leptospira Detection in the Laboratory.'

Health Cycle 1 Recipients