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Published on: June 29, 2020
Development of a cost-effective method for capripoxvirus genotyping using snapback primer and dsDNA intercalating dye
Esayas Gelaye1, Charles Euloge Lamien, Roland Silber
1Animal Production and Health Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Vienna, Austria ; Institute of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria ; Research and Diagnostic Laboratories, National Veterinary Institute, Debre Zeit, Ethiopia.
A new, cost-effective molecular assay accurately genotypes sheep pox virus (SPPV), goat pox virus (GTPV), and lumpy skin disease virus (LSDV). This rapid method aids in controlling capripox diseases by enabling swift identification of the specific virus causing outbreaks.
Area of Science:
- Virology
- Molecular Biology
- Veterinary Diagnostics
Background:
- Sheep pox virus (SPPV), goat pox virus (GTPV), and lumpy skin disease virus (LSDV) are closely related capripoxviruses (CaPVs) causing significant diseases in livestock.
- Complex epidemiology and co-existence of CaPVs in some regions complicate accurate outbreak identification and control.
- Accurate and rapid genotyping is crucial for effective disease management and vaccine selection.
Purpose of the Study:
- To design and evaluate a novel, low-cost molecular assay for simultaneous detection and genotyping of CaPVs.
- To assess the analytical performance, including sensitivity and specificity, of the developed genotyping method.
Main Methods:
- Development of a molecular assay utilizing unlabelled snapback primers and EvaGreen dye for CaPV genotyping.
- Genotyping based on melting temperature analysis of primer-formed hairpins and full-length amplicons.
- Validation using 63 field samples, differentiating between SPPV, GTPV, and LSDV.
Main Results:
- The assay achieved 100% sensitivity and specificity in simultaneously detecting and genotyping CaPVs.
- Successfully genotyped 14 SPPVs, 25 GTPVs, and 24 LSDVs from the tested samples.
- The method is pathogen-specific, cross-platform compatible, and cost-effective, avoiding fluorescent probes and specialized software.
Conclusions:
- The developed molecular assay provides a rapid, specific, and cost-effective solution for CaPV genotyping.
- This assay is suitable for resource-limited diagnostic and research laboratories, facilitating early detection.
- The method will significantly contribute to epidemiological studies and informed control strategies for capripox diseases.
