Improved LNA probe-based assay for the detection of African and South  American yellow fever virus strains.
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.
Weidmann
Manfred
M
University Medical Center, Department of Virology, Kreuzbergring 57, 37075 Göttingen, Germany. mweidma@gwdg.de
Faye
Ousmane
O
Faye
Oumar
O
Kranaster
Ramon
R
Marx
Andreas
A
Nunes
Marcio R T
MR
Vasconcelos
Pedro F C
PF
Hufert
Frank T
FT
Sall
Amadou A
AA
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*
D015345:Oligonucleotide Probes