摘要
目的:建立单克隆抗体(McAb)夹心ELISA法,用于检测SARS病毒(SARS-CoV)抗原。方法:用间接夹心ELISA法筛选捕捉和标记用单克隆抗体的组合,采用过碘酸钠法标记辣根过氧化物酶(HRP),优化后用于检测SARS-CoV。结果:从12株抗SARS-CoV鼠单克隆抗体中筛选出2A3/1C5组合用于捕捉SARS-CoV,1A5/1B4组合标记HRP作为指示抗体。优化后2A3/1C5的最适工作浓度为1∶4000,HRP-1A5/1B4的最适工作浓度为1∶2000。本方法检测SARS-CoV的敏感度为105pfu/mL。结论:单克隆抗体夹心ELISA法可特异性检测SARS-CoV抗原。
Objective: To establish an assay for SARS-CoV by McAbs sandwich ELISA method. Methods: The McAbs for this assay were selected by indirect sandwich ELISA and the McAbs were labeled by HRP. After optimization, McAbs sandwich ELISA method was used for detection of SARS-CoV. Results: Combined 2A3/1C5 and 1A5/1B4 were selected from a panel of twelve McAbs established by immunizing BALB/c mice with inactivated BJ01 strain. The working concentration of the capture McAbs 2A3/1C5 was 1:4 000 and the HRP-1A2/1B4 was 1:2 000. The sensitivity of this assay was 105 pfu/mL. Conclusion: McAbs sandwich ELISA assay could be used specifically for detection of SARS-CoV.
出处
《生物技术通讯》
CAS
2006年第1期34-36,共3页
Letters in Biotechnology