Rapid molecular detection and genotyping of West Nile Virus lineages 1 and 2 by real time PCR and melting curve analysis

J Virol Methods. 2014 Oct:207:54-9. doi: 10.1016/j.jviromet.2014.06.020. Epub 2014 Jun 30.

Abstract

Following its spread in the USA, West Nile Virus (WNV) has reemerged in the Mediterranean basin with a renewed pathogenicity. The introduction of WNV lineage 2 in Europe and its co-circulation with lineage 1 has resulted in a continuously changing epidemiological scenario, highlighting the importance of differential detection of the two lineages. The paper describes a new real-time PCR method for the detection and genotyping of the two main lineages of WNV. The method requires a single pair of primers and probes and is based on the analysis of highly conserved consensus sequences detected in the 5' terminus of the viral genome.

Keywords: Flavivirus; Melting curve analysis; Molecular detection; Virus genotyping; WNV.

MeSH terms

  • DNA Primers / genetics
  • Europe / epidemiology
  • Genome, Viral
  • Genotyping Techniques / methods*
  • Oligonucleotide Probes / genetics
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Transition Temperature*
  • United States / epidemiology
  • West Nile Fever / epidemiology
  • West Nile Fever / virology*
  • West Nile virus / classification*
  • West Nile virus / genetics
  • West Nile virus / isolation & purification*

Substances

  • DNA Primers
  • Oligonucleotide Probes
  • RNA, Viral