Melioidosis, caused by Burkholderia pseudomallei, is a serious infectious disease common in tropical regions but often underdiagnosed and underreported. Existing detection methods are either time-consuming, expensive, or require complex laboratory equipment, making them unsuitable for rapid diagnosis in the field or in low-resource settings. There is an urgent need for a fast, accurate, and cost-effective detection method that can be used at the point of care to enable timely diagnosis, treatment, and control of disease outbreaks.

The project offers a rapid and cost-effective solution through the development of a multiplex quantitative Loop-Mediated Isothermal Amplification (m-qLAMP) assay using dual-function oligonucleotides. These oligonucleotides serve both as primers and signal reporters, enabling real-time monitoring of DNA amplification. The test operates under isothermal conditions, eliminating the need for complex thermal cyclers. It can be used with portable devices like AmpliFire or integrated into existing real-time detection systems. The solution delivers results in under one hour, with high specificity and sensitivity, making it suitable for point-of-care diagnostics in both clinical and field environments. This approach ensures faster decision-making and better disease management, especially in regions where melioidosis is prevalent.

The key innovation of this project lies in the use of dual-function oligonucleotides that act both as primers for DNA amplification and as real-time fluorescent reporters. This dual functionality simplifies the assay design and improves detection efficiency by reducing the number of components needed. The technology is applied in a multiplex quantitative Loop-Mediated Isothermal Amplification (m-qLAMP) format, which enables rapid, accurate detection of Burkholderia pseudomallei under constant temperature conditions, without the need for complex thermal cycling equipment. Unlike traditional PCR methods, this assay allows real-time monitoring of amplification while maintaining high specificity and sensitivity. It is also adaptable to both new portable devices like AmpliFire and existing lab-based real-time detection systems, offering flexibility, cost-effectiveness, and broader usability across different settings.