A novel HRM assay for the simultaneous detection and differentiation of eight poxviruses of medical and veterinary importance View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-12

AUTHORS

Esayas Gelaye, Lukas Mach, Jolanta Kolodziejek, Reingard Grabherr, Angelika Loitsch, Jenna E. Achenbach, Norbert Nowotny, Adama Diallo, Charles Euloge Lamien

ABSTRACT

Poxviruses belonging to the Orthopoxvirus, Capripoxvirus and Parapoxvirus genera share common host species and create a challenge for diagnosis. Here, we developed a novel multiplex PCR method for the simultaneous detection and differentiation of eight poxviruses, belonging to three genera: cowpox virus (CPXV) and camelpox virus (CMLV) [genus Orthopoxvirus]; goatpox virus (GTPV), sheeppox virus (SPPV) and lumpy skin disease virus (LSDV) [genus Capripoxvirus]; orf virus (ORFV), pseudocowpox virus (PCPV) and bovine papular stomatitis virus (BPSV) [genus Parapoxvirus]. The assay is based on high-resolution melting curve analysis (HRMCA) of PCR amplicons produced using genus specific primer pairs and dsDNA binding dye. Differences in fragment size and GC content were used as discriminating power. The assay generated three well separated melting regions for each genus and provided additional intra-genus genotyping allowing the differentiation of the eight poxviruses based on amplicon melting temperature. Out of 271 poxviral DNA samples tested: seven CPXV, 25 CMLV, 42 GTPV, 20 SPPV, 120 LSDV, 33 ORFV, 20 PCPV and two BPSV were detected; two samples presented co-infection with CMLV and PCPV. The assay provides a rapid, sensitive, specific and cost-effective method for the detection of pox diseases in a broad range of animal species and humans. More... »

PAGES

42892

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep42892

DOI

http://dx.doi.org/10.1038/srep42892

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1083849617

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/28216667


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