Please use this identifier to cite or link to this item:http://hdl.handle.net/20.500.12105/6814
A step forward in molecular diagnostics of lyssaviruses:results of a ring trial among European laboratories
Fischer, Melina | Wernike, Kerstin | Freuling, Conrad M | Müller, Thomas | Aylan, Orhan | Brochier, Bernard | Cliquet, Florence | Vazquez-Moron, Sonia ISCIII | Hostnik, Peter | Huovilainen, Anita | Isaksson, Mats | Kooi, Engbert A | Mooney, Jean | Turcitu, Mihai | Rasmussen, Thomas B | Revilla-Fernández, Sandra | Smreczak, Marcin | Fooks, Anthony R | Marston, Denise A | Beer, Martin | Hoffmann, Bernd
PLoS One. 2013;8(3):e58372
Rabies is a lethal and notifiable zoonotic disease for which diagnostics have to meet the highest standards. In recent years, an evolution was especially seen in molecular diagnostics with a wide variety of different detection methods published. Therefore, a first international ring trial specifically designed on the use of reverse transcription polymerase chain reaction (RT-PCR) for detection of lyssavirus genomic RNA was organized. The trial focussed on assessment and comparison of the performance of conventional and real-time assays. In total, 16 European laboratories participated. All participants were asked to investigate a panel of defined lyssavirus RNAs, consisting of Rabies virus (RABV) and European bat lyssavirus 1 and 2 (EBLV-1 and -2) RNA samples, with systems available in their laboratory. The ring trial allowed the important conclusion that conventional RT-PCR assays were really robust assays tested with a high concordance between different laboratories and assays. The real-time RT-PCR system by Wakeley et al. (2005) in combination with an intercalating dye, and the combined version by Hoffmann and co-workers (2010) showed good sensitivity for the detection of all RABV samples included in this test panel. Furthermore, all used EBLV-specific assays, real-time RT-PCRs as well as conventional RT-PCR systems, were shown to be suitable for a reliable detection of EBLVs. It has to be mentioned that differences were seen in the performance between both the individual RT-PCR systems and the laboratories. Laboratories which used more than one molecular assay for testing the sample panel always concluded a correct sample result. Due to the markedly high genetic diversity of lyssaviruses, the application of different assays in diagnostics is needed to achieve a maximum of diagnostic accuracy. To improve the knowledge about the diagnostic performance proficiency testing at an international level is recommended before using lyssavirus molecular diagnostics e.g. for confirmatory testing.
Animals | Europe | Female | Humans | Lyssavirus | Male | RNA, Viral | Reverse Transcriptase Polymerase Chain Reaction | Sensitivity and Specificity | Rhabdoviridae Infections
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