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 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).