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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

Shannon L. Emery*, Dean D. Erdman*Comments to Author , Michael D. Bowen*, Bruce R. Newton*, Jonas M. Winchell*, Richard F. Meyer*, Suxiang Tong*, Byron T. Cook*, Brian P. Holloway*, Karen A. McCaustland*, Paul A. Rota*, Bettina Bankamp*, Luis E. Lowe*, Tom G. Ksiazek*, William J. Bellini*, and Larry J. Anderson*
Author affiliations: *Centers for Disease Control and Prevention, Atlanta, Georgia, USA

Main Article

Table 3

Efficiency of real-time PCR assaysa

Mean CTb values at estimated RNA transcript copy number
SARS1
7.5 x 100
7.5 x 101
7.5 x 102
7.5 x 103
7.5 x 104
7.5 x 105
7.5 x 106
Slopec
Efficiency (%)d
RNA transcript alone
Neg
38.65±1.48
34.25±0.57
31.1±0.14
27.5
24.2
20.55±0.07
–3.55
91.1
RNA transcript + extracte
Neg
38.05±0.92
34.85±0.21
31.55±0.07
27.75±0.07
24.4
20.6
–3.49
93.3
SARS2
2 x 100
2 x 101
2 x 102
2 x 103
2 x 104
2 x 105
2 x 106


RNA transcript alone
35.4±0.57
32.1±0.14
29.45±0.64
26.15±0.07
22.9±0.14
19.4
16.35±0.07
–3.21
104.9
RNA transcript + extract
Neg
34.55±1.91
29.2±0.28
26.2
23.1
19.6±0.14
16.6
–3.48
93.9
SARS3









RNA transcript alone
39.3
36.2±0.42
32.8
29.1±0.14
25.9
22.15±0.07
19.2
–3.39
97.1
RNA transcript + extract 40.3 36.2±0.28 33.4±0.28 29.9±0.21 26.05±0.07 22.55±0.21 19.65±0.21 –3.42 96.1

aPCR, polymerase chain reaction; CT, threshold cycle number.
bValues shown are mean of triplicate samples ± standard deviations.
cSlope determined from the formula: Y = Y intercept – slope log10. Slopes calculated for SARS1 (7.5 x 106 to 7.5 x 101); SARS2 (2 x 106 to 2 x 101); SARS3 (2 x 106 to 2 x 100).
dEfficiency = [10(–1/slope)] – 1.
eReactions performed in presence of pooled total nucleic acid extract from 20 human respiratory specimens.

Main Article

Page created: January 27, 2011
Page updated: January 27, 2011
Page reviewed: January 27, 2011
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