Isolation and Molecular characterization of Contagious Ecthyma (ORF) Virus from Small Ruminants and Human in Egypt

Abstract

Isolation and molecular characterization of ORF virus provides high sensitivity methods for accurate and rapid diagnosis for ORF virus infection in sheep, goat and human in Egypt. Thirty five (35) skin scabs samples (15) from sheep and 15 from goat and 5 biopsy skin samples from human hands) and one hundred and sixteen (116) serum samples were collected from (48 sheep, 29 goats and 39 human) at Beni Suef Governorate, Egypt during the period from July to August 2013. All biopsy samples from human and animals were prepared and inoculated on chorio-allantoic membranes (CAM) of embryonated chicken eggs (ECE) for virus isolation, positive sample caused pock lesions in CAM. The isolated viruses were identified and characterized by Enzyme linked immunesorbent assay (ELISA), Fluorescent antibody technique (FAT), electron microscopy (E/M) and polymerase chain reaction (PCR). They gave specific green fluorescence by FAT, Micrograph showed ovoid shape particles 290-300×160 nm in diameter very closely similar to references ORF virus by using electron microscopy (E/M). Molecular characterization of isolated viruses by PCR with using (B2L gene) fragments approximately 592 bp which typical as reference ORF virus strains. Detection of ORF virus antibodies in the serum samples by protein A ELISA were (10.26%, 31.03% and 37.5%) by IFAT were (7.69%, 20.69% and 25%) and by AGPT were (2.5%, 17.24% and 14.5%) there was significant difference (p<0.05) between all tests used for F virus infection in human or animals (goat and sheep) at Beni-suef Governorate, Egypt respectively. It was concluded that the PCR and protein A ELISA proved to be more rapid simple and sensitive for detection of ORF virus infection in human and animals.