Citation: ZHOU Dun-jin, TANG Shao—wen, WAN G Bin, YAN G Hua-fen, DIAO Ping, XU Qing, LIU Bin, YANG Ji—hong*. One-step Duplex RT-PCR Assay for Detection SARS Associated Coronavirus .VIROLOGICA SINICA, 2003, 18(3) : 232-236.

One-step Duplex RT-PCR Assay for Detection SARS Associated Coronavirus

  • Available online: 18 June 2003
  • According to PCR primers for SARS developed by WHO network laboratories we synthesize 4 primer pairs: BNIin-s/as、BNIout-s/as、SARl-s/as、COR1/COR2 and establish assays of one-step RT-PCR for detection of SARS associated coronavirus. Specific DNA fragments as predicted were produced from specimen W20 by the assays using all 4 primer pairs, while one specific DNA fragment was produced from specimen W1 using primer pair BNIin-s/as. The PCR products from W20 were sequenced and the sequence showed high identity to that of all SARS associated coronavirus in GenBank, which showed that PCR products amplified from W1 and W20 by our assays was specific for SARS associated coronavirus. Based on the one-step RT-PCR we developed four one-step duplex RT-PCR for amplifying two fragment in one reaction by combination of 4 primer pairs above. And the assays of one-step duplex RT-PCR have higher specificity and were reliable for detection of SARS associated coronavirus.

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    One-step Duplex RT-PCR Assay for Detection SARS Associated Coronavirus

    • 1. Wuhan Centersfor Diseases Prevention and Control,Wuhan 430022,China
    • 2. The Key Laboratory ofMolecular Virology,Wuhan Insititute of Virology,The Chinese Academy ofSciences,Wuhan 430071,China

    Abstract: According to PCR primers for SARS developed by WHO network laboratories we synthesize 4 primer pairs: BNIin-s/as、BNIout-s/as、SARl-s/as、COR1/COR2 and establish assays of one-step RT-PCR for detection of SARS associated coronavirus. Specific DNA fragments as predicted were produced from specimen W20 by the assays using all 4 primer pairs, while one specific DNA fragment was produced from specimen W1 using primer pair BNIin-s/as. The PCR products from W20 were sequenced and the sequence showed high identity to that of all SARS associated coronavirus in GenBank, which showed that PCR products amplified from W1 and W20 by our assays was specific for SARS associated coronavirus. Based on the one-step RT-PCR we developed four one-step duplex RT-PCR for amplifying two fragment in one reaction by combination of 4 primer pairs above. And the assays of one-step duplex RT-PCR have higher specificity and were reliable for detection of SARS associated coronavirus.

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