H, A., Z, H., R, H., A, S., M, R., A, A., N, A., Α, A., A, A., S, M. (2008). APPLICATION OF REAL TIME PCR FOR IDENTIFICATION OF SHEEP POXVIRUS, CAMEL POXVIRUS AND CONTAGIOUS ECTHYMA VIRUS ISOLATES. test, 56(2), 37-46. doi: 10.21608/vmjg.2008.378910
Abdel Baky, M H; Hassan, K Z; Habashi, A R; Saad, M A; Rabei M; Abd El-Rahim, I A; AI - Hafufi, A N; AI- Essa, Α Α; AI-Sukayran A; Mazloum, K S. "APPLICATION OF REAL TIME PCR FOR IDENTIFICATION OF SHEEP POXVIRUS, CAMEL POXVIRUS AND CONTAGIOUS ECTHYMA VIRUS ISOLATES". test, 56, 2, 2008, 37-46. doi: 10.21608/vmjg.2008.378910
H, A., Z, H., R, H., A, S., M, R., A, A., N, A., Α, A., A, A., S, M. (2008). 'APPLICATION OF REAL TIME PCR FOR IDENTIFICATION OF SHEEP POXVIRUS, CAMEL POXVIRUS AND CONTAGIOUS ECTHYMA VIRUS ISOLATES', test, 56(2), pp. 37-46. doi: 10.21608/vmjg.2008.378910
H, A., Z, H., R, H., A, S., M, R., A, A., N, A., Α, A., A, A., S, M. APPLICATION OF REAL TIME PCR FOR IDENTIFICATION OF SHEEP POXVIRUS, CAMEL POXVIRUS AND CONTAGIOUS ECTHYMA VIRUS ISOLATES. test, 2008; 56(2): 37-46. doi: 10.21608/vmjg.2008.378910

APPLICATION OF REAL TIME PCR FOR IDENTIFICATION OF SHEEP POXVIRUS, CAMEL POXVIRUS AND CONTAGIOUS ECTHYMA VIRUS ISOLATES

Veterinary Medical Journal (Giza)
Volume 56, Issue 2, April 2008, Pages 37-46    PDF (3.56 M)
Document Type: Original Article
DOI: 10.21608/vmjg.2008.378910
Authors
Abdel Baky, M H* 1; Hassan, K Z2; Habashi, A R3; Saad, M A1; Rabei M4; Abd El-Rahim, I A4; AI - Hafufi, A N4; AI- Essa, Α Α4; AI-Sukayran A5; Mazloum, K S5
1Central Laboratory for Evaluation of Veterinary Biologics, Cairo
2Veterinary Serum and Vaccine Research Institute Abbassia, Cairo
3Dept. of Animal Resource, Ministry of Agriculture, KSA
4Dept. of Veterinary Laboratories. Minstry of Agriculture, KSA
5National Research Center of Agriculture and Animal Resource, KSA
Abstract
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 ñ 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 vi- ruses, and Saudi field isolates of contagious ecthyma (Orf) virus. Capri ñ 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.
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