Development and approbation of primers for identification of lumpy skin desease virus by qPCR method
Journal Title: Бюлетень "Ветеринарна біотехнологія" - Year 2018, Vol 32, Issue
Abstract
Introduction. Lumpy Skin Disease (LSD) is a highly contagious transboundary viral disease of cattle characterized by the formation of necrotizing skin nodules. The OIE requires notification of this disease due to significant economic losses during the outbreak. As the numbers of LSD outbreaks are increasing in the world, including Russia that shares borders with Ukraine, a threat of pathogen introduction into the country is high. The goal of the work was to design primers and a fluorescence probe for LSD virus detection and their approbation with qPCR method. Materials and methods. The specific primers and a fluorescence probe to the part of the viral genome which encoding the core DNA binding phosphoprotein (GenBank: KH 764645) were selected. The developed primers provided amplification of the viral genome fragment with length 151 bp. The Capripoxvirus DNA samples, provided by Dr. Hoffman (FLI, Germany) were used for approbation of developed primers. Both in-house PCR-mix (Thermo Fisher Scientific reagents) and commercial qScript™ XLT (Quanta) were used. Amplification was conducted using Roto-Gene Q. Results of research and discussion. It was confirmed that all Capripoxvirus agents could be detected using the developed primers. The study shown that the results obtained using the in-house PCR-mix were similar to the commercial qScript™ XLT (Quanta). Using the in-house PCR-mix potentially can reduce the test’s cost. It was confirmed 100% specificity and high sensitivity of the developed primers on 18 LSD virus and 14 goat poxvirus DNA samples obtained from experimentally infected cows and goats. Conclusions and prospects for further research. The developed primers can be recommended to use in the veterinary medicine labs for diagnosis of both LSD virus and other Capripoxvirus after the appropriate validation will be done.
Authors and Affiliations
L. Ishchenko, Anna Kovalenko, Larysa Muzykina, Svetlana Mandygra, Ihor Halka, Serghiy Nychyk, V. Spyrydonov
Keywords
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- EP ID EP392515
- DOI 10.31073/vet_biotech32(2)-23
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