Development and Proof-of-Concept Analytical Validation of a HotStart Loop-Mediated Isothermal Amplification (LAMP) Assay Targeting the UL1 Gene for Macacine alphaherpesvirus 1 (McHV-1)

Abstract

Simple Summary Monkey B Virus is a zoonotic pathogen that naturally exists in macaque monkeys and poses a potential risk of fatal infection to humans upon transmission through bites or scratches. Currently, B virus detection requires expensive machines and well-trained technicians in a laboratory. Our method uses a constant warm temperature and a special chemical reaction that produces a clear color change that can be seen with the naked eye—from deep violet to bright yellow-green—when the virus is present. In our experiments using purified target DNA and spiked saliva specimens, this new test detected the viral target within 15 min and was as accurate as the much slower and more complex lab methods. Because this test is quick, easy to use, and does not rely on expensive instruments, it holds promise for future deployment in monkey breeding facilities and resource-limited regions. However, because this study was conducted using laboratory-prepared samples rather than naturally infected specimens, additional testing on real clinical samples will be needed before this method can be used for infection diagnosis.