Preliminary Study on Detecting the SARS-CoV Specific Target cDNA Fragments by Multiplex PCR

Chen, Wenbing; Li, Shousong; Shao, Biying; Zheng, Teng; Jiang, Shuxun; Huang, Xiaorong; Cai, Kaizhen; Zhang, Zhideng

Researchers have made great advancement on the study of preventing, diagnosing and treating the SARS disease, as well as genomic sequencing and detection technology of SARS-CoV (http://www.cas.ac.cn/html/Dir/2003/05/03/ 1847.htm; http://tech.sina.com.cn/ae/2003-05-30/ 0740192503.shtml). However, up to now, only single PCR and ELISA (enzyme-linked immunosorbent assay) have been applied to diagnose SARS-CoV. It has not been reported that multiplex PCR was used to detect SARS-CoV. By amplifying more than two * Corresponding author. E-mail: [e-mail] target fragments in one tube, multiplex PCR can effectively avoid false positive phenomena. Meanwhile, it is low cost and time-saving for detection. So far, multiplex PCR has been used to detect transgenic products (1 ) and many viruses, such as smallpox virus (http://www.who.int/emc/pdfs/smallpoxenglish.pdf) and newcastle disease virus (2-3). Since we were not able to get SARS-CoV sample, and neither could possess a P3 laboratory, we synthesized artificially the target cDNA fragments of SARS-CoV according to the genome sequence of SARS-CoV in GenBank submitted by The Chinese University of Hong Kong, and used it as simulated positive samples. Five primers recommended by World Health Organization (WHO) were used to amplify the fragments by single PCR and multiplex PCR (http://www.who.int/csr/sars/primers/en/print.html).