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

Jessica Coertse,Louis H Nel,Wanda Markotter,Jacqueline Weyer,Negin P Martin

doi:10.1371/journal.pone.0219292

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.

Highlights

Rabies is a fatal viral infection that causes progressive encephalitis in all mammals and is caused by 16 officially recognized viral species [1,2]
As rabies is 100% preventable the United Against Rabies collaboration; consisting of the World Health Organization (WHO), the World Organisation for Animal Health (OIE), the Food and Agriculture Organization of the United Nations (FAO) and the Global Alliance for Rabies Control (GARC); have compiled a global strategic plan in order to eliminate dog-mediated human rabies by 2030 [5] that emphasizes the need for accurate data
A panel of brain material (n = 109) collected from naturally infected animals [8,9,26,27,28,29] that tested positive with the fluorescent antibody test (FAT) [30] and the direct, rapid immunohistochemical test (DRIT) [8] and four negative samples were included for diagnostic evaluation of the RT-Recombinase polymerase amplification (RPA)

Summary

Introduction

Rabies is a fatal viral infection that causes progressive encephalitis in all mammals and is caused by 16 officially recognized viral species [1,2]. It is estimated that approximately 59 000 human deaths occur each year due to rabies and that over 99% of these are caused by rabies virus (RABV) transmitted by domestic dogs [3]. RABV is established worldwide in various hosts and can be divided into two phylogenic groups i.e. bat-related RABV and dog-related RABV. The latter group can be further divided into several major clades i.e. the cosmopolitan, Africa 2, Africa 3, Arctic-related, Asian and Indian subcontinent clades [4].

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