title: A step forward in molecular diagnostics of lyssaviruses--results of a ring trial among European laboratories.
abstract: 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.
authors.affiliation: Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
authors.forename: Melina
authors.initials: M
authors.lastname: Fischer
authors.forename: Kerstin
authors.initials: K
authors.lastname: Wernike
authors.forename: Conrad M
authors.initials: CM
authors.lastname: Freuling
authors.forename: Thomas
authors.initials: T
authors.lastname: Müller
authors.forename: Orhan
authors.initials: O
authors.lastname: Aylan
authors.forename: Bernard
authors.initials: B
authors.lastname: Brochier
authors.forename: Florence
authors.initials: F
authors.lastname: Cliquet
authors.forename: Sonia
authors.initials: S
authors.lastname: Vázquez-Morón
authors.forename: Peter
authors.initials: P
authors.lastname: Hostnik
authors.forename: Anita
authors.initials: A
authors.lastname: Huovilainen
authors.forename: Mats
authors.initials: M
authors.lastname: Isaksson
authors.forename: Engbert A
authors.initials: EA
authors.lastname: Kooi
authors.forename: Jean
authors.initials: J
authors.lastname: Mooney
authors.forename: Mihai
authors.initials: M
authors.lastname: Turcitu
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Rasmussen
authors.forename: Sandra
authors.initials: S
authors.lastname: Revilla-Fernández
authors.forename: Marcin
authors.initials: M
authors.lastname: Smreczak
authors.forename: Anthony R
authors.initials: AR
authors.lastname: Fooks
authors.forename: Denise A
authors.initials: DA
authors.lastname: Marston
authors.forename: Martin
authors.initials: M
authors.lastname: Beer
authors.forename: Bernd
authors.initials: B
authors.lastname: Hoffmann
mesh_terms: D000818:Animals; D005060:Europe; D005260:Female; D006801:Humans; D018114:Lyssavirus / Q000235:genetics*; D008297:Male; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D018353:Rhabdoviridae Infections / Q000175:diagnosis / Q000235:genetics / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral