Thesis (M.Sc.) - Cairo University - Faculty of Veterinary Medicine - Department of Virology

Viral respiratory diseases are frequently reported in equine species affecting equine industry and cause huge economic losses. In the present study, we developed a multiplex assay for the simultaneous detection of the main viruses that cause respiratory diseases in equine species, Equine herpesvirus type 1(EHV-1) and 4 (EHV-4) and Equine influenza virus (EIV). The primers and probes amplified only the targeted viruses and there were no inter- assay cross amplifications or non-specific interactions. The multiplex assay efficiencies were 92.5%, 97% and 90% and the monoplex efficiencies were 97.4%, 98.2% and 90.7% for EHV-1, EHV-4 and EIV, respectively. The R square values (Rsq) in both forms were greater than 0.990. The performance of the assay was evaluated by analysing 152 different clinical samples from clinically infected horses. EHV-1 DNA was detected as the single causative agent in 12 samples, EHV-4 DNA in 9 samples and both EHV-1 and EHV-4 were detected in 4 samples. EIV RNA was not detected during this study. The accuracy of the assay was confirmed by comparing these results with those obtained from analysing the same samples using commercial rPCR and rRT- PCR diagnostic kits. This multiplex assay is proven to be a sensitive, specific, accurate and cost- effective method for the detection of the target viruses whether they occur as a single agent or as a part of co-infections

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