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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
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