Application of real time - PCR for identification of sheep poxvirus, camel poxvirus and contagious ecthyma virus isolates

Recently, attention has been directed toward the application of Real time - PCR assays as a rapid and accurate tools for identification of Capripox, Parapox and Orthopox viruses that cause devastating diseases in farm animals in the Kingdom of Saudi Arabia. SYBR Green [Real time n PCR assays with primer pairs; Capri - Ks.1 of Capripoxviruses, 045 Orf of Parapoxviruses and Q Orf of Pan-parapoxviruses were adopted on a panel of Saudi field isolates and reference strains of sheep and goat poxviruses, camel poxvirus, reference strains of Lumpy skin disease and vaccinia viruses, and Saudi field isolates of contagious ecthyma [Orf] virus. Capri n KS.1 primer set succeeded to amplify all test DNAs of sheep and goat pox, Camel pox, Lumpy skin disease and vaccinia viruses. With melting curve analysis, temperature of melting [Tm] scored by sheep and goat poxvirus, camel pox virus isolates and their reference strains were relatively identical [between 81.1 and 81.8OC], while Tm scored by Lumpy skin disease virus and vaccinia virus were 82.1 and 83.2OC respectively. No amplification was detected from DNAs of Orf virus isolates tested by Capri - KS.1 primers. However, the Orf virus isolates were fairly equivalent amplified with both of 045 Orf primer set [mean Tm score 92.5°C] and Q Orf primer set [mean Tm score 84°C], as well as with TaqMan Real time - PCR in the presence of TaqMan probe and Q Orf primer set. Otherwise, occasional nonspecific amplification of some isolates and strains of Capripox and Orthopox viruses were detected by too late cycle of amplification [> 35 cycle] with 045 Orf and Q Orf primer sets

Isolation and standardization of camelpox virus from naturally infected cases in central region of Saudi Arabia 2004 [with 8 photos and one table]

Camelpox virus in skin lesion scrapings; collected from camels severely affected with clinical signs of camelpox; suspecion was detected by Indirect Fluorescent Antibody [IFA] test using standard anti Jouf-78 strain camelpox virus rabbit antiserum. The causative agent was isolated and purified throughout five successive passages on vero cell cultures using the highest positive dilutions. The virus isolate showed identical Cytopathogenic Effects [CPE] in inoculated vero cell cultures between the 1[st] and 5[th] passages. The isolate was standardized as a virulent camelpox virus by application of IFA, virus neutralization and pathogenicity tests, and designated as Saudi Arabia, Uniza 2004 strain

Immunogenicity of camelpox virus, jouf -78 vaccine - strain in boska t rabbits and guinea pigs [with 2 tables]

Attention was directed to the lack of small laboratory animals [Rabbits and Guinea Pigs] to replace the camels in evaluating the immunogenlcity of the attenuated tissue culture vaccine of camlepox virus, Jouf-78 strain, and in preparation of monospecific antiserum against camelpox virus. The obtained results proved that rabbits reacted immunologically to the inoculated vaccine, and monospecific antiserum against camelpox virus, Jouf-78 strain was successfully prepared in rabbits and guinea pigs