ABSTRACT: To establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR.
Primers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested.
The positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160).
The simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 07/2005; 19(2):176-8.