Reverse transcription recombinase polymerase amplification assay for rapid detection of canine associated rabies virus in Africa

Jessica Coertse; Jacqueline Weyer; Louis H Nel; Wanda Markotter

doi:10.1371/journal.pone.0219292

Reverse transcription recombinase polymerase amplification assay for rapid detection of canine associated rabies virus in Africa

PLoS One. 2019 Jul 5;14(7):e0219292. doi: 10.1371/journal.pone.0219292. eCollection 2019.

Authors

Jessica Coertse¹, Jacqueline Weyer^{1

2}, Louis H Nel³, Wanda Markotter¹

Affiliations

¹ Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.
² The Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases, a Division of the National Health Laboratory Services, Sandringham, South Africa.
³ Centre for Viral Zoonoses, Department of Biochemistry, Genetics and Microbiology, Faculty of Natural and Agricultural Sciences, University of Pretoria, Pretoria, South Africa.

Abstract

Rabies is a neglected disease mostly affecting the developing world. Accurate and reliable diagnostic and surveillance data forms the foundation for the formulation and monitoring of control strategies. Although various sensitive and specific tests are available for detection of rabies virus, implementation of these tests in low-resource settings are challenging and remains limited. In this study, we describe the developed of a reverse transcription recombinase polymerase amplification assay for the detection of rabies virus. The analytical sensitivity of this assay was determined to be 562 RNA copies and was performed in 20 minutes. The diagnostic sensitivity of the RT-RPA was 100% for detection of rabies virus in field samples. In conclusion, the RT-RPA assay allowed for very quick and sensitive detection of rabies virus and could be adapted for use in low-source settings.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Africa / epidemiology
Animals
Biological Assay
Dogs / virology
Nucleic Acid Amplification Techniques / methods
Rabies / diagnosis*
Rabies / veterinary
Rabies virus / genetics*
Rabies virus / pathogenicity
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction / methods*
Sensitivity and Specificity

Grants and funding

This work is based on research supported by the South African Research Chair initiative of the Department of Science and Technology and National Research Foundation of South Africa (Grant No 98339, WM) and the Poliomyelitis Research Foundation (Grant No 16/27, JW).