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Methods Collections

Current Research Methods in Rabies Diagnosis, Prevention, Treatment, and Control

Collection Overview

Rabies is an acute progressive encephalitis caused by a lyssavirus. The case fatality of this zoonosis is the highest for any infectious disease. Although laboratory techniques on rabies have progressed greatly over the last century, typically such information falls in disparate locations and is often unavailable for easy access by those with the greatest need. Hence, the opportunity for a visual, methodological catalog is critical for basic research in pathobiology, immunology and molecular bio Show More

Abstracts

Novel procedure of rabies test for animals by detecting viral antigens stably in Merkel cells at the follicle-sinus complexes of muzzle skins

Nozomi Shiwa*1, Kazunori Kimitsuki2, Daria L. Manalo3, Akira Noguchi4, Satoshi Inoue*4, Chun-Ho Park1
1Kitasato University, 2Oita University, 3Research Institute for Tropical Medicine, 4National Institute of Infectious Diseases

 Rabies is a zoonotic disease caused by lyssaviruses which are transmitted to humans via rabid animals and causes an acute encephalomyelitis after a variable incubation period, and also is the highest case fatality rate of any currently recognized infectious disease. Effective surveillance and monitoring of rabies susceptible animals is a key activity to the assessment of exposed persons. However, the dissection of the head for rabies testing in the laboratory is still laborious with a high Show More

Use of mobile technology in the enhanced implementation of mass dog vaccination interventions

Andy Gibson*1, Gowri Yale2, Ryan Wallace3, Frederic Lohr1, Ian Handel4, Stella Mazeri1, Berend Bronsvoort4, Richard Mellanby*4, Luke Gamble*2
1Mission Rabies, University of Edinburgh, 2Mission Rabies, 3Centres for Disease Control and Prevention, 4University of Edinburgh

Many high income nations have successfully eliminated rabies through annual mass dog vaccination campaigns implemented comprehensively across the country. This requires expertise in project planning, cartography and logistics, the coordination of hundreds of vaccination teams and capacity to aggregate, review and analyse large data sets. There are few low and middle income countries that have demonstrated that this can be achieved at the national scale, let alone refined and sustained to achieve Show More

IN-VITRO TESTS FOR RABIES VACCINE POTENCY TESTING

Noël TORDO*1, Corinne JALLET2
1Antiviral Strategy Unit, Institut Pasteur, Paris, France; Institut Pasteur de Guinée, Conakry, Guinea, 2Antiviral Strategy Unit, Institut Pasteur, Paris, France

he growing global concern for animal welfare is encouraging manufacturers and National Control Laboratories (OMCLs) in the “3Rs strategy” for the “Replacement, Reduction and Refinement” of laboratory animal testing. Development of in vitro approaches are recommended at the WHO and European levels as alternatives to the NIH test for evaluating rabies vaccine potency. At the surface of the rabies virus (RABV) particle, trimers of glycoprotein constitute t Show More

The Rapid Fluorescent Focus Inhibition Test and Adaptions

Susan Moore*1
1Kansas State University, Rabies Laboratory, Manhattan, Kansas 66502 USA

Abstract: The Rapid Fluorescent Focus Inhibition Test (RFFIT) has been in use since the first publication of the assay in 1973, quickly becoming the gold standard for measurement of rabies virus neutralizing antibodies (RVNA). Because the test system mimics the virus-antibody-cell interactions that determine the function of the RVNA (prevention of infection), it is the best ass Show More

Iophenoxic Acid as a biological marker for oral rabies vaccination in the small Indian mongoose

Are Berentsen*1, Robert Sugihara2, Cynthia Payne2, Israel Leinbach2, Steven Volker1, Ad Vos3, Steffen Ortmann3, Amy Gilbert1
1USDA APHIS WS National Wildlife Research Center, 4101 LaPorte Ave., Fort Collins, CO 80521, 2USDA APHIS WS National Wildlife Research Center, 210 Amauulu Rd., Hilo, HI, 96720, 3IDT Biologika GmbH, Am Pharmapark, 06861 Dessau-Rosslau, Germany

The small Indian mongoose (Herpestes auropunctatus) is a reservoir of rabies virus (RABV) in parts of the Caribbean, such as Puerto Rico and comprises over 70% of animal rabies cases reported annually. The control of RABV circulation in wildlife reservoirs is typically accomplished by a strategy of oral rabies vaccination (ORV). Currently no wildlife ORV program exists in Puerto Rico. Research into oral rabies vaccines and Show More

Quantification of rabies virus in different brain structures of livestock

Elizabeth Loza-Rubio*1, Edith Rojas-Anaya1, Daniela Anaya-Razo2, Isabel Bárcenas-Reyes3
1Centro Nacional de Investigación Disciplinaria en Microbiología, INIFAP, 2Universidad del Valle de México, 3Facultad de Ciencias Naturales-Universidad Autónoma de Querétaro

Bovine Paralytic Rabies (BPR) is of great economic importance throughout Latin America. This viral encephalitis produces significant losses to the livestock industry and poses a major zoonotic risk. For rapid and sensitive diagnosis, selection of the best CNS sample is critical. The objective of our work was to utilize a laboratory protocol to quantify the relative number of rabies virus (RV) particles detected in different anatomical portions of the bovin Show More

Pan-lyssavirus SYBR real time RT-PCR for rabies diagnosis

Denise Marston*1, Daisy Jennings1, Nikki MacLaren1, Tony Fooks1, Ashley Banyard1, Lorraine McElhinney*1
1Wildlife Zoonoses & Vector-Borne Diseases Research Group, Animal and Plant Health Agency, New Haw, Addlestone, Surrey, KT15 3NB, United Kingdom

Molecular assays are rapid, sensitive and specific, and have become central to diagnosing rabies. PCR based assays have been utilised for decades to detect rabies, but have only recently been accepted by the OIE as a method to confirm rabies infection. Real-time RT-PCR assays provide real-time data, and are closed-tube systems, minimising the risk of contamination during set up. SYBR real-time RT-PCR assays do not require Show More

Evaluation of a commercial blocking ELISA for the detection of rabies virus antibodies in North American wildlife sera

Amy Gilbert*1, Heather Sullivan1, Charles Rupprecht2, Susan Moore3
1USDA APHIS WS National Wildlife Research Center, 2LYSSA LLC, 3Kansas State University Rabies Laboratory

Rabies lyssavirus (RV) causes an important global zoonosis with an estimated 59,000 human deaths annually. The majority of human cases are transmitted by domestic dogs. Human fatalities within North America are infrequent, yet RV circulates naturally and independently in multiple bat and carnivore reservoirs, resulting in human exposures and prophylaxis. Testing for RV antibodies (RVA) is a key diagnostic activity, widely recognized as a measure of resistance to lethal i Show More

Development and validation of the pan-lyssavirus nested RT-PCR for diagnosis of rabies

Ye Feng*1, Changchun Tu*2, Yuyang Wang1, Weidi Xu1, Huancheng Guo1, Wenjie Gong1, Biao He1
1Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, 21 Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences 2 Jiangsu Co-innovation Centre for Prevention and Control of Important Animal Infectious Diseases and Zoonoses

The pan-lyssavirus nested RT-PCR (RT-nPCR) has been developed to amplify a part of the N gene of lyssaviruses and verified as a highly specific diagnostic tool for detection of all 16 confirmed lyssavirus species classified by ICTV and 2 tentative novel lyssavirus species (Kotalahti bat lyssavirus and Taiwan bat lyssavirus). The validation using 9624 brain specimens collected from different animal species with multiple geographical Show More

Lateral flow test for simple animal rabies diagnosis in resource-limited settings.

Stephanie Mauti*1, Monique Léchenne2, Service Naïssengar3, Abdallah Traoré4, Morgane Gourlaouen5, Paola De Benedictis5, Hervé Bourhy1, Jakob Zinsstag2, Laurent Dacheux*1
1Institut Pasteur, Unit Lyssavirus Dynamics and Host Adaptation, National Reference Center for Rabies and WHO Collaborating center for reference and Research on Rabies, Paris, France, 2Swiss Tropical and Public Health Institute, Basel, Switzerland; University of Basel, Basel, Switzerland, 3Institut de Recherche en Elevage pour le Developpment, Farcha, N’Djamena, Chad, 4Laboratoire Central Vétérinaire, Km 8, Route de Koulikoro, BP 2295, Bamako, Mali, 5FAO Reference Centre for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Italy

High functioning continuous rabies surveillance systems are crucial to provide reliable data and increase the political commitment necessary for successful sustainable disease control. To date, animals suspected as rabid must have a post-mortem confirmation using either classical or molecular laboratory methods. This necessitates prompt submission of diagnostic samples to a functioning and accessible laboratory. However, in most endemic areas, animal rabies diagnosi Show More

High-Resolution 3D Imaging of Rabies Virus Infection in Solvent-Cleared Brain Tissue

Luca Zaeck*1, Madlin Potratz1, Stefan Finke*1
1Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Molecular Virology and Cell Biology, Greifswald – Insel Riems, Germany

Visualization of infection events in tissues and organs by immunolabeling is a key method in modern infection biology. The ability to observe and study the distribution, tropism, and abundance of pathogens inside of organ samples provides pivotal data on disease development and progression. Using conventional microscopy methods, immunolabeling is mostly restricte to thin sections obtained from paraffin-embedded or frozen samples. Because of the limited 2D image plane provided Show More

Laboratory standard operating procedure of lyssavirus surveillance on bat (Chiroptera) population in Taiwan

Wei-Cheng Hsu*1, Shu-Chia Hu*1, Yang-Chang Tu1, Jen-Chieh Chang1, Kwok-Rong Tsai1, Fan Lee1
1Animal Health Research Institute, Council of Agriculture, Executive Yuan, Taiwan

Viruses within the genus Lyssavirus are zoonotic pathogens and at least seven lyssavirus species are are associated with human cases. Because bats are the natural reservoirs of most lyssaviruses, to understand the ecology of these viruses in bats in Taiwan, a lyssavirus surveillance program of bats has been conducted since 2008. For occupational safety concerns, all laboratory workers handle bat specimens only after receiving pre-exposure rabies prophylaxis (PrEP). All procedures should be perfo Show More

Rabies necropsy techniques on small and large specimens

Jodie Jarvis*1, Kathleen Brown1, Patrick Fitzgerald1, April Davis1
1New York State Department of Health, Rabies Laboratory

Safe and efficient necropsy techniques are essential qualities for all rabiesdiagnostic laboratories. Determining the necessary personal protective equipment (PPE) requires evaluation of the necropsy space, instruments in use, and type of specimens that may be submitted. The New York State Department of Health receives between 6,000 and 8,000 specimens annually ranging from juvenile bats to intact submissions from draft horses and adult cattle. A Show More

A Protocol for Enhanced Rabies Surveillance Using a Direct Rapid Immunohistochemical Test

Erin Patrick*1, Brian Bjorklund2, Jordona Kirby3, Kathleen Nelson4, Richard Chipman4, Charles Rupprecht5
1USDA, APHIS, Wildlife Services, Knoxville, TN, 2USDA, APHIS, Wildlife Services, Sutton, MA, 3USDA, APHIS, Wildlife Services, Milton, FL, USA, 4USDA, APHIS, Wildlife Services, Concord, NH, USA, 5LYSSA LLC, Atlanta, GA, USA

Laboratory-based surveillance is integral for rabies prevention, control, and management efforts. While the direct fluorescent antibody test (DFA) is the gold standard for rabies diagnosis, there is an ongoing need to validate additional diagnostic techniques to improve rabies surveillance, particularly in developing countries. Here, we present a standard protocol for a direct rapidimmunohistochemical test  Show More


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