title: A high-resolution melting (HRM) assay for the differentiation between Israeli field and Neethling vaccine lumpy skin disease viruses.
abstract: Lumpy skin disease (LSD) is a constant threat to the Middle East including the State of Israel. During vaccination programs it is essential for veterinary services and farmers to be able to distinguish between animals affected by the cattle-borne virulent viruses and vaccinated animals, subsequently affected by the vaccine strain. This study describes an improved high resolution-melting (HRM) test that exploits a 27 base pair (bp) fragment of the LSDV126 extracellular enveloped virion (EEV) gene that is present in field viruses but is absent from the Neethling vaccine strain. This difference leads to ∼0.5 °C melting point change in the HRM assay, when testing the quantitative PCR (qPCR) products generated from the virulent field viruses compared to the attenuated vaccine. By exploiting this difference, it could be shown using the newly developed HRM assay that virus isolated from vaccinated cattle that developed disease symptoms behave similarly to vaccine virus control, indicating that the vaccine virus can induce disease symptoms. This assay is not only in full agreement with the previously published PCR gradient and restriction fragment length polymorphism (RFLP) tests but it is faster with, fewer steps, cheaper and dependable.
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Sophia
authors.initials: S
authors.lastname: Menasherow
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Oran
authors.initials: O
authors.lastname: Erster
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Marisol
authors.initials: M
authors.lastname: Rubinstein-Giuni
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Anita
authors.initials: A
authors.lastname: Kovtunenko
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Eyngor
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Boris
authors.initials: B
authors.lastname: Gelman
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Khinich
authors.affiliation: Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel. Electronic address: yehudastram@gmail.com.
authors.forename: Yehuda
authors.initials: Y
authors.lastname: Stram
mesh_terms: D000818:Animals; D002417:Cattle; D003937:Diagnosis, Differential; D008166:Lumpy Skin Disease / Q000175:diagnosis* / Q000821:virology*; D008167:Lumpy skin disease virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D008877:Middle East; D025202:Molecular Diagnostic Techniques / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D044366:Transition Temperature; D014730:Veterinary Medicine / Q000379:methods*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000009:adverse effects*
chemical_list: D014765:Viral Vaccines
keywords: EEV gene; HRM; High-resolution melting; LSDV126 gene; Lumpy Skin Disease Virus