Real-Time RT-PCR for the Detection of Lyssavirus Species

Deubelbeiss, Andrea; Zahno, Marie-Luise; Zanoni, Manuela; Brügger, Dorothy; Zanoni, Reto Giacomo (2014). Real-Time RT-PCR for the Detection of Lyssavirus Species. Journal of Veterinary Medicine, 2014, pp. 1-12. Hindawi 10.1155/2014/476091

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The causative agents of rabies are single-stranded, negative-sense RNA viruses in the genus Lyssavirus of Rhabdoviridae, consisting of twelve classified and three as yet unclassified species including classical rabies virus (RABV). Highly neurotropic RABV causes rapidly progressive encephalomyelitis with nearly invariable fatal outcome. Rapid and reliable diagnosis of rabies is highly relevant for public and veterinary health. Due to growing variety of the genus Lyssavirus observed, the development of suitable molecular assays for diagnosis and differentiation is challenging. This work focused on the establishment of a suitable real-time RT-PCR technique for rabies diagnosis as a complement to fluorescent antibody test and rabies tissue culture infection test as gold standard for diagnosis and confirmation. The real-time RT-PCR was adapted with the goal to detect the whole spectrum of lyssavirus species, for nine of which synthesized DNA fragments were used. For the detection of species, seven probes were developed. Serial dilutions of the rabies virus strain CVS-11 showed a 100-fold higher sensitivity of real-time PCR compared to heminested RT-PCR. Using a panel of thirty-one lyssaviruses representing four species, the suitability of the protocol could be shown. Phylogenetic analysis of the sequences obtained by heminested PCR allowed correct classification of all viruses used.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

UniBE Contributor:

Deubelbeiss, Andrea; Zahno, Marie-Luise; Zanoni, Manuela; Brügger, Dorothy and Zanoni, Reto Giacomo

Subjects:

600 Technology > 630 Agriculture

ISSN:

2356-7708

Publisher:

Hindawi

Language:

English

Submitter:

Susanne Portner

Date Deposited:

24 Apr 2015 12:04

Last Modified:

31 Oct 2018 10:42

Publisher DOI:

10.1155/2014/476091

BORIS DOI:

10.7892/boris.67222

URI:

https://boris.unibe.ch/id/eprint/67222

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