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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 1

Primers and probes used for real-time RT-PCR assaysa

Assay ID Primer/ probe Sequence (5′>3′) Genomic region Locationb
Primary diagnostic assay




SARS1
F
CAT GTG TGG CGG CTC ACT ATA T
RNA polymerase
15370-15392

R
GAC ACT ATT AGC ATA AGC AGT TGT AGC A

15422-15449

P
TTA AAC CAG GTG GAA CAT CAT CCG GTG

15395-15420
SARS2
F
GGA GCC TTG AAT ACA CCC AAA G
Nucleocapsid
28531-28552

R
GCA CGG TGG CAG CAT TG

28581-28597

P
CCA CAT TGG CAC CCG CAA TCC

28559-28574
SARS3
F
CAA ACA TTG GCC GCA AAT T
Nucleocapsid
29016-29034

R
CAA TGC GTG ACA TTC CAA AGA

29063-29083

P
CAC AAT TTG CTC CAA GTG CCT CTG CA

29036-29061
To confirm positive results




N3
F
GAA GTA CCA TCT GGG GCT GAG
Nucleocapsid
28432-28452

R
CCG AAG AGC TAC CCG ACG

28383-28400

P
CTC TTT CAT TTT GCC GTC ACC ACC AC

28406-28431
3′NTR
F
AGC TCT CCC TAG CAT TAT TCA CTG
3′ nontranslated region
29619-29642

R
CAC CAC ATT TTC ATC GAG GC

29576-29595

P
TAC CCT CGA TCG TAC TCC GCG T

29597-29618
M
F
TGT AGG CAC TGA TTC AGG TTT TG
Membrane protein
26951-26973

R
CGG CGT GGT CTG TAT TTA ATT TA

27005-27027
P CTG CAT ACA ACC GCT ACC GTA TTG GAA 26974-27000

aRT-PCR, reverse transcription–polymerase chain reaction; F, forward primer; R, reverse primer; P, probe.
bLocation based on the severe acute respiratory syndrome–associated coronavirus, Urbani strain (GenBank accession number AY278741).

Main Article

Page created: January 27, 2011
Page updated: January 27, 2011
Page reviewed: January 27, 2011
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
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