Smartphone-Linked and Electricity-Free Platforms for Rapid Colorimetric Molecular Detection of Poultry Respiratory Viruses at the Point of Need.

Abstract

Efficient control measures for respiratory diseases in humans and farm animals require accurate, specific, and rapid diagnostics. Traditional PCR-based molecular diagnostics are restricted to centralized laboratories, which results in significant, potentially catastrophic delays in test results. A case in point is the recent avian flu outbreak, which has culled more than 280 million poultry birds worldwide (over 157 million in the USA alone) since 2022; has spread to other farm animals, such as cattle; has further heightened the risk of a human pandemic; and threatens food security. To enable molecular diagnosis of bird respiratory diseases at the point of need, we employ loop-mediated isothermal amplification (LAMP) in two platforms: (A) portable devices linked to a smartphone and (B) an inexpensive, disposable, electricity-free, instrument-free device with closed-tube, colorimetric detection that can be produced with minimal resources. Smartphone integration offers an unexplored opportunity for spatiotemporal disease mapping, equipping policymakers with critical data for outbreak control. Our assays demonstrated 100% sensitivity and specificity compared to the gold standard, lab-based, quantitative PCR (qPCR). We tested contrived samples of the avian flu H5N1 virus, laryngotracheitis virus (ILTV), and infectious bronchitis virus (IBV) spiked into clinical samples, achieving a detection sensitivity adequate for early infection diagnosis in under 45 min. The test is simple, requires minimal training, and can be performed without refrigeration, making it well-suited for resource-limited settings.