الفهرس 
Development of ELISA kit for detection of equine herpes viruses causing abortion among equines in Egypt
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Abstract
In Egypt, The losses caused by EHVs infections in equine studs during last years increased more than ever, especially with the increase in Arabian horses{u2019} trade and traveling of Egyptian horses to participate in international championships all over the world. Economic losses had reached millions of Egyptian pounds, especially for valuable Arabian breeds, indeed the suspect was EHV-1; however the epidemic had not subsided with the strict vaccination regimen for it. This initiated the present design of a protocol for isolation of EHV from field specimens to stand on the real cause, then prepare a seed isolate to develop a homemade ELISA kit for EHV antigen detection that is not offered commercially to assure rapid, accurate alternative with cost efficiency. A total of 287 samples were collected including 200 serum samples, 12 Nasal and 18 uterine swabs from suspected cases and 57 tissue samples from aborted feti and their dams. Rapid screening was performed using PCR on 87 samples of tissues and swabs. Four samples were found Positive for the presence of EHV-4 specific N.A, sequencing and phylogenetic based analysis were done. Viral isolation was carried out from PCR positive samples on MDBK cell line, followed by identification methods; AGID, IFAT, VNT and PCR. Virus titration was done on the sixth passage which revealed a titer of 10⁶·⁷⁵. Steps for antigen preparation were made starting with concentration using PEG-6000 followed by dialysis as initial purification ended with differential centrifugation using high speed centrifugation for final purification, then whole cell antigen preparation was done using sonication