Rapid point of care diagnosis of equine Hendra Virus using LAMP technology

Summary

Hendra virus (HeV) is a highly lethal zoonotic paramyxovirus. In the first 20 years after discovery, 52 HeV incidents have occurred involving 94 equine and 7 human cases (Smith et al 2016). The most recent case of HeV was reported in June 2019 occurring in a brood mare in the Hunter Valley, New South Wales. All human fatalities recorded thus far highlight the occupational nature of zoonotic infection. Currently, qRTPCR (realtime quantitative reverse transcription PCR) assay is the gold standard for detection of HeV, with Biosecurity Queensland (BQ) analyses on average 1,500 diagnostic samples annually (5.7 daily). Processing and transport of samples results in long delays of results leading to considerable stress on industry stakeholders and prevents timely treatment of these almost 6 horses a day in QLD. Initial development and preliminary investigation of a rapid loopmediated isothermal amplification (LAMP) test to detect HeV on specialised instrumentation in close collaboration with BQ has shown considerable promise. Due to its simplicity, robustness, and low cost, LAMP can be used as a rapid assay at the point of care by clinicians, making it directly applicable for use in the equine industry. Five equine facilities in HeV endemic regions, GeneWorks, BQ and the Equine Veterinarians Australia support in principle and financially this research. This research aims to achieve 3 goals; 1) optimisation of the LAMP HeV test; 2) field testing by veterinarians of LAMP with mock samples in parallel with qRTPCR in the laboratory; and 3) develop an interactive computer based biosecurity work flow system integrated with LAMP testing to maximise performance and safety.

Program

Thoroughbred Horses

Research Organisation

The University of Queensland

Objective Summary

The major objectives of this project are: 1) Optimise the HeV LAMP test to perform at a level that will allow it to be deployed to protect human health and improve horse welfare. 2) Undertake field testing to evaluate practicality and functionality when operated by veterinarians, and compare with parallel results from qRTPCR submissions. Evaluate specificity and limit of detection correlation between diagnostic laboratory results and results obtained by LAMP inclinic or within the field by veterinarians. 3) Develop, in collaboration with GeneWorks, the EVA and BQ an interactive computer based biosecurity work flow system LAMP testing to maximise performance and safety without compromising assay performance. 4) Evaluate the fitness for purpose of the LAMP assay through a rigorous assay validation in comparison with the gold standard The expected outcomes are: 1) Assay sensitivity optimisation of the LAMP test to equivalency with qRTPCR viral detection. Initial investigation of the feasibility of the HeV LAMP test has identified several assay optimisation options that will improve limit of detection 2) Implement the LAMP test and associated computer biohazard control system with collaborating equine veterinary professionals in regions with known HeV outbreaks (already identified and engaged) to maximise human safety and accuracy of results in a field setting. This will result in a modular computer controlled approach that will be incorporated into training modules allowing for field application in potentially stressful biohazard locations.

Project Code

PRJ-012093

Project Stage

Current

Project Start Date

Friday, May 15, 2020

Project Completion Date

Tuesday, June 15, 2021

Journal Articles From Project

Not Available

National Priority

An environmentally sustainable Australia

National Priority

Soil, water and managing natural resources

National Priority

HOR-Improve the safety of industry participants and the welfare of horses, and enhance the sustainability of the industry

Contact