Table. Real Time reverse transcription-polymerase chain reaction (RT-PCR) analysis of RNA extracts from 2000 Israel West Nile patient specimens using primer set NY1999-NS5.

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
2.5x106	16.4	1:101	4.5x106	1:101	4.4x105	1	29.3	6.9x102	2.5x106	16.5	cereb.	31.1	1.4x102
2.5x105	20.0	1:102	3.4x105	1:102	4.0x104	2	25.6	7.3x103	2.5x105	20.0	cortex	36.4h	1.9x100
2.5x104	23.6	1:103	3.1x104	1:103	4.3x103	3	30.0	4.6x102	2.5x104	23.1
2.5x103	27.2	1:104	3.1x103	1:104	6.0x102	5	34.8	2.3 x101	2.5x103	26.7
2.5x102	31.1	1:105	3.4x102	1:105	9.8x101				2.5x102	30.3
2.5x101	34.9	1:106	3.1x101	1:10	2.1x100				2.5x101	34.1
2.5x100	36.8h	1:107	3.0x100 h	1:107	n.di				2.5x100	37.3 h
0	>45	0	0	0	0				0	>45

^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 C_T , 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.