Establishment of a fluorescent polymerase chain reaction method for the detection of the SARS-associated coronavirus and its clinical application.

Wu, Xinwei; Cheng, Gang; Di, Biao; Yin, Aihua; He, Yunshao; Wang, Ming; Zhou, Xinyu; He, Lijuan; Luo, Kai; Du, Lin

OBJECTIVE To establish a fluorescent polymerase chain reaction (F-PCR) method for detecting the coronavirus related to severe acute respiratory syndrome (SARS) and to evaluate its value for clinical application. METHODS The primers and the fluorescence-labeled probe were designed and synthesized according to the published sequence of the SARS-associated coronavirus genes. A F-PCR diagnosis kit for detecting the coronavirus was developed, and 115 clinical nasopharyngeal gargling liquid samples were tested. RESULTS The sequence of PCR amplified products completely matched the related sequence of the SARS-associated coronavirus genome. Forty-nine out of 67 samples from identified SARS patients and 8 of 18 samples from persons having close contact with SARS patients showed positive results. All 30 samples from healthy controls were negative. CONCLUSION The F-PCR method established may be a rapid, accurate and efficient way for screening and for the early diagnosis of SARS patients.