Molecular diagnosis of lumpy skin disease by polymerase chain reaction and dna sequencing

Abstract EN

During summer 2005 there was severe outbreak of lumpy skin disease (LSD) in Demiatta, Egypt.

Twenty three samples (18) buffy coats and (5) skin biopsies were collected from infected cows and submitted to laboratory examination for Neethling virusl agent using tissue culture and polymerase chain reaction (PCR).

PCR was superior in detection of Neethling virus (65.2 %) while, tissue culture was lower (34.8 %).

PCR products was used to compare of LSD virus of Ismaillia strain (1989 outbreak), Demiatta strain (2005 outbreak) and sheep pox vaccine, Kenyan strain (SPV) using sequence analysis to determine molecular differences.

Nucleotide sequencing and phylogenetic analysis of 192 bp fragment indicated a close relationship of Capri poxviruses, and there were molecular difference based upon nucleotide sequences with ability to differentiate between sheep pox and LSD.

Ismaillia and Demiatta isolates appeared to be in a separate clad, suggesting that both isolates derived from different origin.

In contrast isolate of Egypt 2005 outbreak is 100 % identical to ELkholy strain (published in gene bank of accession number EU807974, Egypt 2006).