A real time high-resolution melting PCR assay for detection and differentiation among sheep pox virus, goat pox virus, field and vaccine strains of lumpy skin disease virus

Pestova, Yana; Byadovskaya, Olga; Кононов, А. В.; Sprygin, Alexander

doi:10.1016/j.mcp.2018.08.003

Cited by 21 publications

(17 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Lamien et al developed a PCR system for distinction of SPPV and GTPV based on a 21-nucleotide deletion in the SPPV genome compared to GTPV [ 22 ]. In addition, a gel-based duplex PCR assay [ 23 ], a real-time PCR assay containing a snapback primer and a dsDNA intercalating dye [ 24 ] as well as a real-time high-resolution melting PCR assay [ 25 ] are already published.…”

Section: Introductionmentioning

confidence: 99%

Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants

Wolff

King

Moritz

et al. 2020

Viruses

View full text Add to dashboard Cite

Capripox viruses, with their members “lumpy skin disease virus (LSDV)”, “goatpox virus (GTPV)” and “sheeppox virus (SPPV)”, are described as the most serious pox diseases of production animals. A GTPV isolate and a SPPV isolate were sequenced in a combined approach using nanopore MinION sequencing to obtain long reads and Illumina high throughput sequencing for short precise reads to gain full-length high-quality genome sequences. Concomitantly, sheep and goats were inoculated with SPPV and GTPV strains, respectively. During the animal trial, varying infection routes were compared: a combined intravenous and subcutaneous infection, an only intranasal infection, and the contact infection between naïve and inoculated animals. Sheep inoculated with SPPV showed no clinical signs, only a very small number of genome-positive samples and a low-level antibody reaction. In contrast, all GTPV inoculated or in-contact goats developed severe clinical signs with high viral genome loads observed in all tested matrices. Furthermore, seroconversion was detected in nearly all goats and no differences concerning the severity of the disease depending on the inoculation route were observed. Conclusively, the employed SPPV strain has the properties of an attenuated vaccine strain, consistent with the genetic data, whereas the GTPV strain represents a highly virulent field strain.

show abstract

Section: Introductionmentioning

confidence: 99%

Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants

Wolff

King

Moritz

et al. 2020

Viruses

View full text Add to dashboard Cite

show abstract

“…Though several assays are available for CaPV species determination by PCR and real time PCR 18–20,30 , none are intended for differentiation of SPPV vaccines from SPPV field isolates. Various research groups developed assays to differentiate the LSDV Neethling vaccine from LSDV field isolates in cattle 14,15 , and furthermore genotype CaPVs 16 ; however, those assays are unable to differentiate SPPV vaccines from SPPV field isolates. Therefore, it is expected that this HRM assay will be used as a direct means for screening samples from suspected capripox cases collected in unvaccinated flocks, and to undertake epidemiological investigations when a capripox outbreak occurs following vaccination in a small ruminant or cattle herds.…”

Section: Discussionmentioning

confidence: 99%

“…This would help to elucidate the origin of disease in the event of mild or systemic post-vaccination reactions in vaccinated animals 13 . Similar assays have proven to be very useful in cattle to differentiate the LSDV Neethling vaccine from LSDV fields isolates in Israel 14 , Greece 15 and Russia 16 .…”

Section: Introductionmentioning

confidence: 99%

“…With the new generation of dyes and technological improvements in qPCR platforms, high-resolution melting (HRM) curve analysis has become a powerful technique for genotyping microbes, offering the possibility of developing methods other than sequencing for rapid pathogen typing 19,21–24 , including the differentiation of vaccine strains from field isolates 14,16 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

An HRM Assay to Differentiate Sheeppox Virus Vaccine Strains from Sheeppox Virus Field Isolates and other Capripoxvirus Species

Rufael

Settypalli

Berguido

et al. 2019

Sci Rep

View full text Add to dashboard Cite

Sheep poxvirus (SPPV), goat poxvirus (GTPV) and lumpy skin disease virus (LSDV) affect small ruminants and cattle causing sheeppox (SPP), goatpox (GTP) and lumpy skin disease (LSD) respectively. In endemic areas, vaccination with live attenuated vaccines derived from SPPV, GTPV or LSDV provides protection from SPP and GTP. As live poxviruses may cause adverse reactions in vaccinated animals, it is imperative to develop new diagnostic tools for the differentiation of SPPV field strains from attenuated vaccine strains. Within the capripoxvirus (CaPV) homolog of the variola virus B22R gene, we identified a unique region in SPPV vaccines with two deletions of 21 and 27 nucleotides and developed a High-Resolution Melting (HRM)-based assay. The HRM assay produces four distinct melting peaks, enabling the differentiation between SPPV vaccines, SPPV field isolates, GTPV and LSDV. This HRM assay is sensitive, specific, and provides a cost-effective means for the detection and classification of CaPVs and the differentiation of SPPV vaccines from SPPV field isolates.

show abstract

“…Currently there is no DIVA vaccine and companion diagnostic test for capripoxviruses, although there are molecular based methods available to discriminate between vaccine and wild type viruses (142)(143)(144). The development of a DIVA capripoxvirus vaccine and companion diagnostic test is theoretically feasible and technically possible.…”

Section: Recombinant Capripoxvirus Vaccines As Diva Vaccinesmentioning

confidence: 99%

Potential of Using Capripoxvirus Vectored Vaccines Against Arboviruses in Sheep, Goats, and Cattle

Teffera

Babiuk

2019

Front. Vet. Sci.

View full text Add to dashboard Cite

Capripoxviruses represent a genus of the poxviridae family under the subfamily chordopoxviriniae; the genus includes three animal virus species that have a devastating impact on sheep, goats, and cattle in Africa, Asia, and most recently Eastern Europe (1-4). The viruses in the genus are sheeppox virus (SPPV), goatpox virus (GTPV), and lumpy skin disease virus (LSDV) which affect sheep, goats, and cattle, respectively (1, 4). Capripoxviruses share 98% sequence similarity between

show abstract

A real time high-resolution melting PCR assay for detection and differentiation among sheep pox virus, goat pox virus, field and vaccine strains of lumpy skin disease virus

Cited by 21 publications

References 10 publications

Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants

Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants

An HRM Assay to Differentiate Sheeppox Virus Vaccine Strains from Sheeppox Virus Field Isolates and other Capripoxvirus Species

Potential of Using Capripoxvirus Vectored Vaccines Against Arboviruses in Sheep, Goats, and Cattle

Contact Info

Product

Resources

About