Lyssavirus Detection and Typing Using Pyrosequencing

摘要: Rabies is a fatal zoonosis caused by nonsegmented negative-strand RNA virus, namely, rabies virus (RABV). Apart from RABV, at least 10 additional species are known as rabies-related lyssaviruses (RRVs), and some of them responsible for occasional spillovers into humans. More have also been detected recently in different bat ecosystems, thanks to the application molecular diagnostic methods. Due variety members genus Lyssavirus, there necessity develop reliable assay diagnosis able detect differentiate among existing viruses. In present study, pyrosequencing protocol targeting 3' terminus nucleoprotein (N) gene was applied rapid characterization lyssaviruses. Correct identification achieved each sample tested. Results were confirmed those obtained using Sanger sequencing method. A pan-lyssavirus one-step reverse transcription (RT)-PCR developed within framework procedure. The sensitivity (Se) RT-PCR determined vitro-transcribed serial dilutions titrated demonstrated high analytical relative specificity (Sp) (98.94%) (99.71%). To date, this first case which has lyssavirus cheaper approach than one all other protocols typing that we acquainted with. study indicate procedure suitable detection samples both human animal origin.