Development of a Real Time PCR protocol for the diagnosis of Borna disease

. A. CERSINI, A. DAMIANI, G. MANNA, D. AMADDEO , M. SCICLUNA , C. CANTILE, G. AUTORINO

Congreso; X Congress of the Italian Society of Veterinary Diagnosis, SIDILV; 2008

Phylogenetic analysis distributes WNV strains from different origins within four lineages. Diagnosis of WNV infections can be carried out by serological, immunohistochemical, histological and molecular methods. Different real time RT PCR specific for the 3? non coding (NC) genome region, the E and the NS5 genes have been developed. Within a project working on diagnostic tools for viral equine diseases, we developed a real time RT PCR targeting an area of the gene mainly present in NS2a, containing a conserved region of WNV. Our assay resulted more sensitive than the one targeting NC, detecting also a reference strain belonging to the lineage 2 (WNV B956). Specificity was assessed positively by testing the method toward Borna virus (BDV) and Equine herpesvirus type 1 (EHV1), both present in the european equine population. To define the efficiency of the method an mRNA-NS2a was synthetized, which could usefully be employed as a non hazardous positive control in diagnostic routine.