title: Multiplex PCR for rapid diagnosis and differentiation of pox and pox-like diseases in dromedary Camels.
abstract: BACKGROUND
  Pox and pox-like diseases of camels are a group of exanthematous skin conditions that have become increasingly important economically. Three distinct viruses may cause them: camelpox virus (CMLV), camel parapox virus (CPPV) and camelus dromedary papilloma virus (CdPV). These diseases are often difficult to differentiate based on clinical presentation in disease outbreaks. Molecular methods such as PCR targeting species-specific genes have been developed and used to identify these diseases, but not simultaneously in a single tube. Recently, multiplex PCR has gained reputation as a convenient diagnostic method with cost-and timesaving benefits.
  
  
  RESULTS
  In the present communication, we describe the development, optimization and validation of a multiplex PCR assay able to detect simultaneously the genome of the three viruses in one single test allowing for rapid and efficient molecular diagnosis. The assay was developed based on the evaluation and combination of published and new primer sets and was validated with viral genomic DNA extracted from known virus strains (n = 14) and DNA extracted from homogenized clinical skin specimens (n = 86). The assay detects correctly the target pathogens by amplification of targeted genes, even in case of co-infection. The method showed high sensitivity, and the specificity was confirmed by PCR-product sequencing.
  
  
  CONCLUSIONS
  This assay provide rapid, sensitive and specific method for identifying three important viruses in specimens collected from dromedary camels with varying clinical presentations.
authors.affiliation: Camel Research Center, King Faisal University, Al-Ahsa, 31982, Saudi Arabia. abdokhlf@yahoo.co.uk.|Department of Microbiology, Faculty of Veterinary Medicine, University of Khartoum, P.O. Box 32, Shambat, Sudan. abdokhlf@yahoo.co.uk.
authors.forename: Abdelmalik I
authors.initials: AI
authors.lastname: Khalafalla
authors.affiliation: Department of Biochemistry, Physiology and Pharmacology, College of Veterinary Medicine and Animal Resources, King Faisal University, Al-Ahsa, 31982, Saudi Arabia. kbusadah@kfu.edu.sa.
authors.forename: Khalid A
authors.initials: KA
authors.lastname: Al-Busada
authors.affiliation: Central Biotechnology Laboratory, Faculty of Veterinary Medicine and Animal Resources, King Faisal University, Al-Ahsa, 31982, Saudi Arabia. ibsabagh@kfu.edu.sa.|Department of Virology, Cairo University, Giza, 12211, Egypt. ibsabagh@kfu.edu.sa.
authors.forename: Ibrahim M
authors.initials: IM
authors.lastname: El-Sabagh
mesh_terms: D000818:Animals; D002162:Camelus; D003937:Diagnosis, Differential; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D018155:Orthopoxvirus / Q000302:isolation & purification; D027383:Papillomaviridae / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D018157:Parapoxvirus / Q000302:isolation & purification; D011213:Poxviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods*