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

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

*المؤلف المقابل لهذا العمل

نتاج البحث: المساهمة في مجلةمقالمراجعة النظراء

42 اقتباسات (Scopus)

ملخص

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.

اللغة الأصليةEnglish
رقم المقال42892
دوريةScientific Reports
مستوى الصوت7
المعرِّفات الرقمية للأشياء
حالة النشرPublished - فبراير 20 2017
منشور خارجيًانعم

ASJC Scopus subject areas

  • ???subjectarea.asjc.1000???

بصمة

أدرس بدقة موضوعات البحث “A novel HRM assay for the simultaneous detection and differentiation of eight poxviruses of medical and veterinary importance'. فهما يشكلان معًا بصمة فريدة.

قم بذكر هذا