Detection of Crimean–Congo Hemorrhagic Fever, Hanta, and Sandfly Fever Viruses by Real-Time RT-PCR

The development of sensitive and specific nucleic acid diagnostic assays for viral pathogens is essential for proper medical intervention

Sofi M. Ibrahim

2010

Scholarcy highlights

  • The development of sensitive and specific nucleic acid diagnostic assays for viral pathogens is essential for proper medical intervention
  • This chapter describes four fluorescence-based PCR assays to detect the Crimean–Congo Hemorrhagic Fever, Andes, Hantaan, and Sandfly Fever Sicilian Viruses. These assays are based on species-specific hydrolysis probes targeting the nucleocapsid protein gene for CCHFV and SFSV and the glycoprotein gene for ANDV and HANV
  • All four assays were optimized for LightCycler 2.0 or Ruggedized Advanced Pathogen Identification Device
  • The assays were evaluated using the protocols described in the Subheading 3
  • The sensitivity and specificity of the assays were evaluated with test panels that consisted of 20–60 known positive and 30–135 known negative samples, representing 7–34 genetically diverse bacterial and viral species
  • The SFSV assay missed three positive samples and cross-reacted with one of 48 negative samples, resulting in 95% sensitivity at LOD of 5 plaque-forming units and 98% specificity

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