Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group
Abstract Lumpy skin disease (LSD) outbreaks in Southeast and South Asia are attributed to different lineages of LSD virus (LSDV). Variants belonging to the novel recombinant cluster 2.5 circulate in China and Thailand, while a Kenyan sheep and goat pox (KSGP) strain from cluster 1.1 circulates in In...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-09-01
|
| Series: | BMC Research Notes |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13104-023-06502-z |
| _version_ | 1827635750991036416 |
|---|---|
| author | Alexander Sprygin Ali Mazloum Antoinette Van Schalkwyk Alena Krotova Olga Bydovskaya Larisa Prokhvatilova Ilya Chvala |
| author_facet | Alexander Sprygin Ali Mazloum Antoinette Van Schalkwyk Alena Krotova Olga Bydovskaya Larisa Prokhvatilova Ilya Chvala |
| author_sort | Alexander Sprygin |
| collection | DOAJ |
| description | Abstract Lumpy skin disease (LSD) outbreaks in Southeast and South Asia are attributed to different lineages of LSD virus (LSDV). Variants belonging to the novel recombinant cluster 2.5 circulate in China and Thailand, while a Kenyan sheep and goat pox (KSGP) strain from cluster 1.1 circulates in India, Pakistan, and Bangladesh. The clusters representing these circulating strains are vastly different. However, if their distribution encroaches into each other’s ranges, it will be impossible to differentiate between them due to the lack of suitable molecular tools. Thus, fit-for-purpose molecular tools are in demand to effectively and timeously diagnose and investigate the epidemiology of LSDVs in a region. These could significantly contribute to the phylogenetic delineation of LSDVs and the development of preventive measures against transboundary spillovers. This work aimed to develop a real-time polymerase chain reaction assay targeting open reading frame LW032, capable of specifically detecting KSGP-related isolates and recombinant LSDV strains containing the KSGP backbone. The analytical specificity was proven against the widest possible panel of recombinant vaccine-like LSDV strains known to date. The amplification efficiency was 91.08%, and the assay repeatability had a cycle threshold variation of 0.56–1.1 over five repetitions across three runs. This KSGP-specific assay is reliable and fast and is recommended for use in LSDV epidemiological studies where the accurate detection of KSGP genetic signatures is a priority, particularly in regions where KSGP-like and other lineages are circulating. |
| first_indexed | 2024-03-09T15:30:54Z |
| format | Article |
| id | doaj.art-2239d5cb03a84392803a81dc26961f06 |
| institution | Directory Open Access Journal |
| issn | 1756-0500 |
| language | English |
| last_indexed | 2024-03-09T15:30:54Z |
| publishDate | 2023-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Research Notes |
| spelling | doaj.art-2239d5cb03a84392803a81dc26961f062023-11-26T12:14:09ZengBMCBMC Research Notes1756-05002023-09-011611710.1186/s13104-023-06502-zDevelopment and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox groupAlexander Sprygin0Ali Mazloum1Antoinette Van Schalkwyk2Alena Krotova3Olga Bydovskaya4Larisa Prokhvatilova5Ilya Chvala6Federal Center for Animal HealthFederal Center for Animal HealthAgricultural Research Council-Onderstepoort Veterinary InstituteFederal Center for Animal HealthFederal Center for Animal HealthFederal Center for Animal HealthFederal Center for Animal HealthAbstract Lumpy skin disease (LSD) outbreaks in Southeast and South Asia are attributed to different lineages of LSD virus (LSDV). Variants belonging to the novel recombinant cluster 2.5 circulate in China and Thailand, while a Kenyan sheep and goat pox (KSGP) strain from cluster 1.1 circulates in India, Pakistan, and Bangladesh. The clusters representing these circulating strains are vastly different. However, if their distribution encroaches into each other’s ranges, it will be impossible to differentiate between them due to the lack of suitable molecular tools. Thus, fit-for-purpose molecular tools are in demand to effectively and timeously diagnose and investigate the epidemiology of LSDVs in a region. These could significantly contribute to the phylogenetic delineation of LSDVs and the development of preventive measures against transboundary spillovers. This work aimed to develop a real-time polymerase chain reaction assay targeting open reading frame LW032, capable of specifically detecting KSGP-related isolates and recombinant LSDV strains containing the KSGP backbone. The analytical specificity was proven against the widest possible panel of recombinant vaccine-like LSDV strains known to date. The amplification efficiency was 91.08%, and the assay repeatability had a cycle threshold variation of 0.56–1.1 over five repetitions across three runs. This KSGP-specific assay is reliable and fast and is recommended for use in LSDV epidemiological studies where the accurate detection of KSGP genetic signatures is a priority, particularly in regions where KSGP-like and other lineages are circulating.https://doi.org/10.1186/s13104-023-06502-zPolymerase chain reactionKenyan sheep and goat poxVaccineRecombinationReal-timeEpidemiology |
| spellingShingle | Alexander Sprygin Ali Mazloum Antoinette Van Schalkwyk Alena Krotova Olga Bydovskaya Larisa Prokhvatilova Ilya Chvala Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group BMC Research Notes Polymerase chain reaction Kenyan sheep and goat pox Vaccine Recombination Real-time Epidemiology |
| title | Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group |
| title_full | Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group |
| title_fullStr | Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group |
| title_full_unstemmed | Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group |
| title_short | Development and application of a real-time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the Kenyan sheep and goat pox group |
| title_sort | development and application of a real time polymerase chain reaction assay to detect lumpy skin disease virus belonging to the kenyan sheep and goat pox group |
| topic | Polymerase chain reaction Kenyan sheep and goat pox Vaccine Recombination Real-time Epidemiology |
| url | https://doi.org/10.1186/s13104-023-06502-z |
| work_keys_str_mv | AT alexandersprygin developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup AT alimazloum developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup AT antoinettevanschalkwyk developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup AT alenakrotova developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup AT olgabydovskaya developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup AT larisaprokhvatilova developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup AT ilyachvala developmentandapplicationofarealtimepolymerasechainreactionassaytodetectlumpyskindiseasevirusbelongingtothekenyansheepandgoatpoxgroup |