Sensitive and Quantitative Detection of Severe Acute Respiratory Syndrome Coronavirus Infection by Real-Time Nested Polymerase Chain Reaction

• Published in: Clinical infectious diseases 2004-01, Vol.38 (2), p.293-296
• Main Authors: Jiang, Shih Sheng, Chen, Tsan-Chi, Yang, Jyh-Yuan, Hsiung, Chao A., Su, Ih-Jen, Liu, Ying-Lan, Chen, Po-Cheng, Juang, Jyh-Lyh
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Brief Reports; Coronavirus; DNA, Viral - analysis; Human viral diseases; Humans; Infections; Infectious diseases; Medical sciences; Polymerase chain reaction; Polymerase Chain Reaction - methods; Reverse transcriptase polymerase chain reaction; RNA; SARS; SARS coronavirus; SARS virus; SARS Virus - genetics; SARS Virus - isolation & purification; Sensitivity and Specificity; Severe acute respiratory syndrome; Severe Acute Respiratory Syndrome - diagnosis; Severe Acute Respiratory Syndrome - virology; Viral diseases; Viral diseases of the respiratory system and ent viral diseases; Viral RNA; Viruses
• ISSN: 1058-4838; 1537-6591
• DOI: 10.1086/380841

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.