title: Improved LNA probe-based assay for the detection of African and South  American yellow fever virus strains.
abstract: BACKGROUND
  Real-time assays for Yellow fever virus (YFV) would help to improve acute diagnostics in outbreak investigations.
  
  
  OBJECTIVE
  To develop a real-time assay for YFV able to detect African and South American strains.
  
  
  METHODS
  Three short probe (14-18 nt) formats were compared and a plasmid-transcribed RNA standard was used to test the performance of the assays. Additionally the new TaqM1 enzyme was tested.
  
  
  RESULTS
  A locked nucleotide probe (LNA probe) performed best with an analytical sensitivity of 10 RNA molecules detected. 44 African and 10 South American strains were detectable. One South American strain from 1984 had a one-nucleotide deviation in the hybridisation sequence for which the LNA probe had to be adapted. Comparison of enzymes revealed that not all enzymes are suitable for LNA probes.
  
  
  CONCLUSIONS
  The developed LNA probe based YFV real-time PCR performed best in an enzyme mix and less efficient using multifunctional enzymes.
authors.affiliation: University Medical Center, Department of Virology, Kreuzbergring 57, 37075 Göttingen, Germany. mweidma@gwdg.de
authors.forename: Manfred
authors.initials: M
authors.lastname: Weidmann
authors.forename: Ousmane
authors.initials: O
authors.lastname: Faye
authors.forename: Oumar
authors.initials: O
authors.lastname: Faye
authors.forename: Ramon
authors.initials: R
authors.lastname: Kranaster
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
authors.forename: Marcio R T
authors.initials: MR
authors.lastname: Nunes
authors.forename: Pedro F C
authors.initials: PF
authors.lastname: Vasconcelos
authors.forename: Frank T
authors.initials: FT
authors.lastname: Hufert
authors.forename: Amadou A
authors.initials: AA
authors.lastname: Sall
mesh_terms: D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*; D015004:Yellow Fever / Q000175:diagnosis* / Q000821:virology; D015005:Yellow fever virus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D015345:Oligonucleotide Probes