[Gene detection of severe acute respiratory syndrome-related coronavirus].

Wu, Bing-quan; Zhong, Hao-hao; Gao, Jian-ping; Liu, Shu-ping; Heng, Wan-jie; E, Wen; Gu, Jiang

OBJECTIVE To develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood. METHODS A pair of primers and a probe (molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank (G130027616). RESULTS In the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process. The 145 bp fragment of PCR product was further confirmed by conventional PCR assay and proved by DNA sequencing to be identical to the target sequence to which the probe was hybridized. CONCLUSION This assay has a broad application for clinical diagnosis and surveillance investigation.