Sensitive and quantitative detection of severe acute respiratory syndrome coronavirus infection by real-time nested polymerase chain reaction.
A quantitative, real-time, nested polymerase chain reaction (PCR) method, combining the high sensitivity of nested PCR with time-saving real-time instrumentation, was developed for large-scale screening for severe acute coronavirus (SARS) coronavirus. Forty-six clinical specimens were analyzed by this method, and results were compared with those obtained by conventional, single-round, real-time reverse-transcriptase PCR (RT-PCR) performed in parallel. Of the 17 positive results, 2 identified by our method were not detected by single-round, real-time RT-PCR, which suggests that real-time nested PCR has the potential for increased sensitivity, leading to earlier detection of SARS.
Jiang
Shih Sheng
SS
Division of Molecular and Genomic Medicine, National Health Research Institutes, Taipei, Taiwan.
Chen
Tsan-Chi
TC
Yang
Jyh-Yuan
JY
Hsiung
Chao A
CA
Su
Ih-Jen
IJ
Liu
Ying-Lan
YL
Chen
Po-Cheng
PC
Juang
Jyh-Lyh
JL
D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D016133: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
D004279:DNA, Viral