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Research article - Peer-reviewed, 2011

Pan-serotypic detection of foot-and-mouth disease virus using a minor groove binder probe reverse transcription polymerase chain reaction assay

McKillen, John; McMenamy, Michael; Reid, Scott M.; Duffy, Catherine; Hjertner, Bernt; King, Donald P.; Belák, Sándor; Welsh, Michael; Allan, Gordon

Abstract

A novel assay for the pan-serotypic detection of foot-and-mouth disease virus (FMDV) was designed using a 5′ conjugated minor groove binder (MGB) probe real-time RT-PCR system. This assay targets the 3D region of the FMDV genome and is capable of detecting 20 copies of a transcribed RNA standard. The linear range of the test was eight logs from 2×101 to 2×108 copies and amplification time was approximately 2h. Using a panel of 83 RNA samples from representative FMDV isolates, the diagnostic sensitivity of this test was shown to be equivalent to a TaqMan real-time RT-PCR that targets the 5′ untranslated region of FMDV. Furthermore, the assay does not detect viruses causing similar clinical diseases in pigs such as swine vesicular disease virus and vesicular stomatitis virus, nor does it detect marine caliciviruses causing vesicular exanthema. The development of this assay provides a useful tool for the differential diagnosis of FMD, potentially for use in statutory or emergency testing programmes, or for detection of FMDV RNA in research applications.

Keywords

Foot-and-mouth disease virus; Real-time PCR; Minor groove binder probe

Published in

Journal of Virological Methods
2011, Volume: 174, number: 1-2, pages: 117-119
Publisher: ELSEVIER SCIENCE BV

      UKÄ Subject classification

      Veterinary Science
      Animal and Dairy Science

      Publication identifier

      DOI: https://doi.org/10.1016/j.jviromet.2011.03.008

      Permanent link to this page (URI)

      https://res.slu.se/id/publ/46572