Volume 8, Number 5—May 2002
Phylogenetic Analysis of a Human Isolate from the 2000 Israel West Nile virus Epidemic
|NS5 standard NY1999||Armored RNA||Armored RNA extract||NY1999 specimens||NS5 standard ISR2000||ISR2000 specimens|
|Amounta||CTb||Dil.c d||Amount||Dil.e||Amountd||Patient no.||CT||Amount||Amountf||CT||Sample||CT||Amountg|
|2.5x100||36.8h||1:107||3.0x100 h||1:107||n.di||2.5x100||37.3 h|
a Plasmid DNA p88-D-21 was quantitated spectrophotometrically, and dilutions containing the indicated copy number of target sequence were added to each polymerase chain reaction (PCR) assay.
b CT , Cycle number at which signal crosses threshold.
c Armored RNA West Nile virus (HNY1999) standard (Ambion, Austin, TX) was diluted 1:10, boiled, reverse transcribed, and then diluted to result in amounts per PCR assay equivalent to the indicated dilution of the stock (5 μL).
d Amount calculated based on calibration curve obtained with NS5 Standard NY1999 (column 1).
e Dilutions of Armored RNA West Nile virus (HNY1999) standard (Ambion) were extracted with TRI-Reagent (Molecular Research Center, Cincinnati, OH) and then subjected to RT-PCR to result in amounts per assay equivalent to the indicated dilution of the stock (5 μL).
f Plasmid DNA pISR-Dfrag-D6 was quantitated spectrophotometrically, and dilutions containing the indicated copy number of target sequence were added to each PCR assay.
g Amount calculated based on calibration curve obtained with NS5 Standard ISR2000 (column 6).
h Poisson effects take place at low template concentration; duplicate assay deviations: 36.2 / 37.4, NY1999; 6.0 x100 / 0, armored RNA; 37.4 / 37.1, ISR2000; and 36.4 / >45, cortex.
i n.d. = not determined.
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