Developing pan lyssavirus beta actin real time RT PCR assays for rabies diagnostics

Rabies is a deadly zoonotic disease caused by rabies virusRABVand other lyssavirusesAnnuallyrabies causes approximatelyhuman deaths worldwideThe only widely accepted diagnostic test is the direct fluorescent antibodyDFAtestWhile rapid and sensitivethe DFA test requires expensivehighquality reagents and equipmentalong with trained diagnosticiansThese factors present economic and logistical barriers that hamper its use in resource limited countriesThe Centers for Disease Control has developed a real time RT PCR based diagnostic assaycalled LNwhich detects allspecies of the Lyssavirus genusNo other assay detects this breadth of lyssavirusesThe assay is rapidsensitiveand easy to perform using standard diagnostic laboratory equipmentcircumventing several limitations of the DFA assayThis proposal aims to optimize LNinto an assay that can be widely adopted by domestic and international labsThe goals of the project are tocombine the LNand beta actin detection assays into a single reactiontest low cost enzymes and reduce reaction volumes to reduce costs anddevelop a dry reagent format to enhance stability of the assay componentsIf successfulthis new low costuserfriendly assay will fill an important gap in the rabies diagnostics field