Characterisation and Next-generation Sequencing Analysis of Unknown Arboviruses
DEFENCE SCIENCE AND TECHNOLOGY ORGANISATION VICTORIA (AUSTRALIA) HUMAN PROTECTION AND PERFORMANCE DIV
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In the last few years there has been an increased incidence of neurological and encephalitic syndromes in Australia that have not been linked to a known pathogen. Viruses from the families Rhabdoviridae and Bunyaviridae are postulated to play a role. This report describes the characterisation of two unknown bunyaviruses, designated Maprik MPKV and Buffalo Creek BCV, and two unknown rhabdoviruses, designated Harrison Dam HARDV and Holmes Jungle HOJV, using techniques such as PCR-select subtraction and next-generation sequencing. Preliminary analysis of the four sequenced viruses has shown that they are all novel, and phylogenetic studies will now be performed to further classify these organisms. In addition, the report details the success in using a novel method to prepare MPKV and BCV RNA for next-generation sequencing. The use of this method allows for an increased level of efficiency when sequencing unknown viruses, and has now been incorporated into our standard viral preparation protocols.