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Animal and Plant Health Agency

2 ARTICLES PUBLISHED IN JoVE

JoVE Methods Collections: Current Research Methods in Rabies Diagnosis, Prevention, Treatment, and Control

Charles E. Rupprecht¹, Wei-Cheng Hsu², Feng Ye³, Megan Golding⁴, Luca Zaeck⁵, Laurent Dacheux⁶, Noel Tordo⁷, Jodie A. Jarvis⁸, Erin Patrick⁹, Are Berentsen¹⁰

¹LYSSA LLC, ²Animal Health Research Institute, Council of Agriculture, Executive Yuan, ³Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, ⁴Wildlife Zoonoses & Vector-Borne Diseases Research Group, Animal and Plant Health Agency, ⁵Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, ⁶Unit Lyssavirus Epidemiology and Neuropathology, National Reference Center for Rabies and WHO Collaborating Center for Reference and Research on Rabies, Institut Pasteur, ⁷Unit Antiviral Strategies, OIE Reference Laboratories for Rift Valley Fever Virus & Crimean Congo Hemorrhagic Fever Virus, Institut Pasteur, Institut Pasteur de Guinea, Conakry, ⁸Rabies Laboratory, Wadsworth Center, New York State Department of Health, ⁹Animal and Plant Health Inspection Service, Wildlife Services, Knoxville, TN, US Department of Agriculture, ¹⁰Animal and Plant Health Inspection Service, Wildlife Services, National Wildlife Research Center, US Department of Agriculture

JoVE Methods Collections: Current Research Methods in Rabies Diagnosis, Prevention, Treatment, and Control

Immunology and Infection

Pan-lyssavirus Real Time RT-PCR for Rabies Diagnosis

Denise A. Marston¹, Daisy L. Jennings¹, Nikki C. MacLaren¹, Daniel Dorey-Robinson¹, Anthony R. Fooks^1,2, Ashley C. Banyard¹, Lorraine M. McElhinney^1,2

¹Wildlife Zoonoses & Vector-Borne Diseases Research Group, Animal and Plant Health Agency, ²Institute of Infection and Global Health, University of Liverpool

This real-time RT-PCR using dsDNA intercalating dye is suitable to diagnose lyssavirus infections. The method begins with RNA extracted from rabies suspected ante-mortem or post-mortem samples, detailing master mix preparation, RNA addition, setup of the real-time machine and correct interpretation of results.