A loop-mediated isothermal amplification assay for rapid and sensitive detection of bovine papular stomatitis virus

J Virol Methods. 2016 Dec:238:42-47. doi: 10.1016/j.jviromet.2016.07.031. Epub 2016 Oct 14.

Abstract

Bovine papular stomatitis virus (BPSV) causes pustular cutaneous disease in cattle worldwide. This paper describes the development of a specific loop-mediated isothermal amplification (LAMP) assay to detect BPSV which did not cross-react with other parapoxviruses. To assess analytical sensitivity of this LAMP assay, DNA was extracted from serially diluted BPSV from which the infectious titer was determined by a novel assay based on calf kidney epithelial cells. The LAMP assay had equivalent analytical sensitivity to quantitative PCR, and could detect as few as 86 copies of viral DNA per reaction. These results suggest that the assay is a specific and sensitive technique to rapidly diagnose bovine papular stomatitis in domestic animals.

Keywords: Bovine papular stomatitis virus; Loop-mediated isothermal amplification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Cattle Diseases / diagnosis*
  • Cattle Diseases / virology
  • DNA Primers / genetics
  • DNA, Viral / analysis
  • Epithelial Cells / virology
  • Limit of Detection
  • Nucleic Acid Amplification Techniques / methods*
  • Parapoxvirus / genetics*
  • Parapoxvirus / isolation & purification
  • Poxviridae Infections / diagnosis
  • Poxviridae Infections / virology
  • RNA, Viral
  • Real-Time Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Temperature

Substances

  • DNA Primers
  • DNA, Viral
  • RNA, Viral