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23

JVS. 2017; 2(1): 92-101


first isolation and identification of EHV-4 causing abortion in Egypt

Ahmed Fawzy Afify, Basem M. Ahmed, Sayed Ahmed Salem, Ahmed Abd El-Ghani El-Sanousi, Mohamed Abd El-Hameed Shalaby.

Abstract
Background: Usually EHV-4 was strictly confined to upper respiratory tract infection with rare incidence of abortion due to rare occurrence of cell associated Viraemia unlike EHV-1 infection, however; suspected herpes viral abortion cases in that outbreak were diagnosed and confirmed EHV-4 cases, Being considered an alteration in pathogenic potentiality of the virus.
Objective: to possess an EHV-4 full identified isolate from field strains to be used as a seed virus in diagnostic purposes later and to update the national monovalent vaccine of EHV-1.
Methods: Full molecular identified two EHV-4 PCR positive samples with a Genbank accession numbers of: KP699582, KX866264 were used in the current study. The samples were obtained during an outbreak of abortion in Arabian horse stud in Egypt during foaling season of 2014-2015. Isolation was carried out on MDBK cell line, identification of EHV-4 isolate was performed using IFAT, AGID and nPCR.
Results: Specific CPE was observed between 4-6th days in first and second passages, time was shortened till development of complete CPE till 2nd day in 5th and 6th passages. EHV-4 isolate was identified by detection of viral antigen with indirect IFAT, AGID tests using reference antisera and finally by nested PCR. The isolate was titrated on MDBK Cell culture revealing a titer of 106.75
Conclusion: EHV-4 can be detected and isolated from abortion cases, also to have an EHV-4 isolate is the first step to follow up the situation of EHV-4 in Egypt as a preliminary step for control of the disease.

Key words: Isolation, MDBK, IFAT, AGID, EHV-4



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