Perspectives on molecular detection methods of lyssaviruses.
Lyssaviruses belong to the family Rhabdoviridae within the order Mononegavirales. Their genomes consist of a negative stranded linear RNA. Although dog transmitted rabies causes the majority of the estimated 55 000 worldwide fatalities per year, bat lyssaviruses have also caused human cases. Based on a broad range of different host species, their geographical distribution and the evolutionary age lyssaviruses display a high degree of genetic variability. Thus the development of molecular diagnostics for the reliable detection and identification of members of the genus Lyssavirus (pan-lyssavirus assay) is a challenge, but is valuable for the fight against rabies worldwide. In this study different published PCR-systems or primers for the detection of a wide range of lyssaviruses were tested as real-time RT-PCRs with different one-step and two-step assays. Comparison of different two-step assays by varying the RT-chemistry revealed differences in sensitivity. Nevertheless, most of the tested one-step systems provided an improved performance, including a reduced assay time and a reduced risk of cross-contaminations, when compared to the optimized two-step assay. Finally, we also provide an overview of additional state-of-the-art molecular methods to detect and differentiate lyssavirus species in general.
Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Greifswald - Insel Riems, Germany.
Melina
M
Fischer
Bernd
B
Hoffmann
Conrad M
CM
Freuling
Thomas
T
Müller
Martin
M
Beer
D000818:Animals; D017931:DNA Primers; D003955:Diagnostic Tests, Routine / Q000592:standards*; D006801:Humans; D018114:Lyssavirus / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis; D011818:Rabies / Q000175:diagnosis* / Q000821:virology*; D011820:Rabies virus / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D012367:RNA, Viral