De Novo synthesis of PCR templates for the development of SARS diagnostic assay

Zhang, Jia; Meng, Bo; Liao, Duanfang; Zhou, Lvyi; Zhang, Xu; Chen, Linling; Guo, Zifen; Peng, Cuiying; Zhu, Bingyang; Lee, Peggy P.; Xu, Xiangmin; Zhou, Tianhong; Deng, Zemin; Hu, Yinghe; Li, Kai

A novel coronavirus was identified as the cause for severe acute respiratory syndrome (SARS). The complete sequence of SARS genome has provided an opportunity for the development of molecular diagnostic assays. To restrain further outbreak of SARS, the World Health Organization has posted several pairs of polymerase chain reaction (PCR) primers for early diagnosis and urged more research to be done on PCR protocols. Here we report a strategy for the de novo synthesis of PCR templates complimentary to the SARS virus genome, which has the advantage of working on PCR templates without concern about viral infection and also has the advantage that it can be used by those who do not have access to the SARS virus. This highly efficient and safe strategy for obtaining SARS gene fragments is useful for the development of PCR assays, as well as for the preparation of reliable positive controls for PCR testing kits.