A one step quantitative RT-PCR for detection of SARS coronavirus with an internal control for PCR inhibitors.
In this study, we report a one step quantitative RT-PCR assay for severe acute respiratory syndrome (SARS) diagnosis. The overall detection rate for clinical samples collected from Days 1 to 9 of disease onset is 86.2% and the specificity of the assay is 100%. To detect false negative results due to PCR inhibitors or faulty RNA extraction, we have further modified this one step RT-PCR assay for simultaneous detection of severe acute respiratory syndrome (SARS) coronavirus (CoV) and 18S ribosomal RNA (rRNA) as an internal positive control.
Poon
Leo L M
LL
Department of Microbiology, Queen Mary Hospital, University of Hong Kong, Pokfulam, Hong Kong SAR, PR China. llmpoon@hkucc.hku.hk
Wong
Bonnie W Y
BW
Chan
Kwok Hung
KH
Leung
Cynthia S W
CS
Yuen
Kwok Yung
KY
Guan
Yi
Y
Peiris
Joseph S M
JS
D004275:DNA, Ribosomal / Q000032:analysis; D006801:Humans; D009305:Nasopharynx / Q000821:virology; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis* / Q000821:virology
D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D012367:RNA, Viral