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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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