Development of a novel Real Time PCR for the detection of the NS2A gene of West Nile Virus

A. CERSINI, I. CIABATTI, A. DAMIANI, G. MANNA, M. SCICLUNA , G. AUTORINO

Congreso; 8th International Congress of Veterinary Virology; 2009

WNV belongs to the family of Flaviviridae, genus Flavivirus and is grouped into the Japanese Encephalitis antigen complex. Four genetic lineages with specific geographic distributions have been described. The Real Time RT-PCRs reported in literature, targeted to the 3? non coding (NC) region, to the E and the NS5 genes, are mainly specific to strains belonging to lineage 1. Recently lineage 2 was detected in both the Mediterranean area as well as in northern Europe, therefore our group proceeded with the development of a novel Real-time RT-PCR for the detection of the aforementioned lineages. Moreover, an in vitro transcribed RNA, specific for target NS2a (mRNA NS2a) has been synthetized to estimate the efficiency of the method and also as a non hazardous positive control in diagnostic routine.