Rapid, Sensitive, LAMP Assay for Monkeypox Virus Clade Differentiation

Orthopox monkeypox (MPXV) has recently emerged as a global viral threat with two distinct clades identified as endemic pathogens. It is estimated that 70% of the world’s population is vulnerable to infection by members of orthopoxviridae.  Rapid identification of MPXV clades is critical, as Clade I infections are more clinically severe than Clade II. Current MPXV differentiation diagnostics either cannot distinguish between the different clades or require additional post-amplification steps, increasing time, cost, and complexity. These limitations reduce the effectiveness of point-of-care testing, especially in outbreak prone and low-resource settings.

 

Researchers at Arizona State University have developed a rapid, sensitive, and specific loop-mediated isothermal amplification (LAMP) assay with specialized primer sets for detecting and differentiating Monkeypox virus clades and Orthopoxvirus lineages. This assay utilizes a single, one-tube amplification reaction with no downstream processing and can be carried out in less than one hour. Designed for point-of-care use, it works directly on crude samples such as skin lesions and bodily fluids, requiring minimal equipment and no sample purification. Additionally, it can be seamlessly integrated into portable microfluidic cartridges for field diagnostics, providing rapid and reliable results validated against conventional qPCR benchmarks.

 

This cost-effective assay represents a novel method for differentiating between highly homologous viral variants for viral surveillance of pathogens in PoC settings.