Volume 10, Number 2—February 2004
THEME ISSUE
2004 SARS Edition
Laboratory Study
Real-Time Reverse Transcription–Polymerase Chain Reaction Assay for SARS-associated Coronavirus
Table 2
Reproducibility of real-time RT-PCR assaysa
RNA transcript copy numberb | ||||||
---|---|---|---|---|---|---|
SARS1 |
7.5 x 101 |
7.5 x 102 |
7.5 x 103 |
7.5 x 104 |
7.5 x 105 |
7.5 x 106 |
CV within assay (%)c |
2.53 |
0.96 |
0.49 |
0.69 |
1.66 |
0.7 |
CV between assays (%)d |
2.39 |
1.09 |
0.82 |
0.64 |
2.1 |
0.79 |
SARS2 |
2.0 x 101 |
2 x 102 |
2 x 103 |
2 x 104 |
2 x 105 |
2 x 106 |
CV within assay (%) |
1.27 |
0.57 |
0.46 |
0.72 |
0.84 |
0.67 |
CV between assays (%) |
1.54 |
1.18 |
0.93 |
1.47 |
1.54 |
1.32 |
SARS3 |
||||||
CV within assay (%) |
0.8 |
0.55 |
0.65 |
0.5 |
0.27 |
1.25 |
CV between assays (%) | 0.94 | 0.64 | 1.07 | 1.13 | 1.24 | 1.65 |
aRT-PCR, reverse transcription–polymerase chain reaction; CV, coefficient of variation.
bTen-fold dilutions of the polymerase and nucleocapsid RNA transcripts; copies per reaction; dilution series thawed on 3 different days and assays performed in triplicate for each dilution.
cDetermined from three replicates within each assay.
dDetermined from three independent assays performed on different days.