A Multiplex RT-PCR Method for the Detection of Reptarenavirus Infection

Author:

Baggio Francesca12,Hetzel Udo1ORCID,Prähauser Barbara12,Dervas Eva1,Michalopoulou Eleni1,Thiele Tanja1,Kipar Anja1ORCID,Hepojoki Jussi13ORCID

Affiliation:

1. The BIBD Group and Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, 8057 Zurich, Switzerland

2. Center for Clinical Studies, Vetsuisse Faculty, University of Zurich, 8057 Zurich, Switzerland

3. Department of Virology, Medicum, Faculty of Medicine, University of Helsinki, 00290 Helsinki, Finland

Abstract

Reptarenaviruses cause Boid Inclusion Body Disease (BIBD), a fatal disease of boid snakes with an economic and ecological impact, as it affects both captive and wild constrictor snakes. The clinical picture of BIBD is highly variable but often only limited. Intracytoplasmic inclusion bodies (IB), which develop in most cell types including blood cells, are the pathognomonic hallmark of BIBD; their detection represents the diagnostic gold standard of the disease. However, IBs are not consistently present in clinically healthy reptarenavirus carriers, which can, if undetected, lead to and maintain the spread of the disease within and between snake populations. Sensitive viral detection tools are required for screening and control purposes; however, the genetic diversity of reptarenaviruses hampers the reverse transcription (RT) PCR-based diagnostics. Here, we describe a multiplex RT-PCR approach for the molecular diagnosis of reptarenavirus infection in blood samples. The method allows the detection of a wide range of reptarenaviruses with the detection limit reaching 40 copies per microliter of blood. Using 245 blood samples with a reference RT-PCR result, we show that the technique performs as well as the segment-specific RT-PCRs in our earlier studies. It can identify virus carriers and serve to limit reptarenavirus spreading in captive snake collections.

Funder

the Sigrid Jusélius Foundation

Publisher

MDPI AG

Subject

Virology,Infectious Diseases

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