Equine herpesvirus-l (EHV-l) is one of the infectious viruses affecting family equidae causing epidemic abortion, neonatal mortality, respiratory diseases and neurological disorders. Due to the economic significant of the disease it is essential to update the circulating types, so this study aimed to isolate the EHV-1 with antigenic and molecular identification of circulating isolates in Giza. A total of 30 tissue samples (aborted fetal tissues, fetal fluids and placenta) were collected from aborted mares of native breeds and different ages during 2018-2020) and 42vaginal and nasal swabs were collected from aborted mares and adult horses of different ages with a history of respiratory manifestations. The samples were prepared and a trial for virus isolation was done on Vero cell culture. Positive samples showing a characteristic cytopathic effect for EHV-1. Identification of positive EHV-1 isolates was done by fluorescence antibody technique (FAT) and polymerase chain reaction (PCR) targeting the glycoprotein B (gpB).The study revealed that 12.5% of the samples were positive for EHV-1. In conclusion, EHV-1infection monitoring was used to investigate and to gain knowledge about natural infection as well as to implement and regulate existing directives concerning clinical diseases. This study recommended that; further molecular characterization will help a better understanding of the virus epidemiology and effective control measures.