基于N蛋白稳定表达细胞株的SARS-CoV间接免疫荧光检测方法的建立

目的:建立基于SARS-CoV N蛋白稳定表达细胞系的间接免疫荧光法,以在普通实验室对SARS-CoV特异抗体进行检测.方法:将含有SARS-CoV N基因的重组质粒通过脂质体转染BHK-21细胞,筛选稳定表达细胞系并制备间接免疫荧光抗原片,以鼠抗SARS血清进行检测,并以其他呼吸道病毒抗体评估其特异性.结果与结论:建立了可检测SARS-CoV抗体的基于SARS-CoV N蛋白稳定表达重组细胞系的间接免疫荧光法,检测其他呼吸道病毒抗体为阴性,特异性良好.该间接免疫荧光法为SARS-CoV抗体的普通实验室检测提供了快速安全的诊断方法.

Immunofluorescence assay for detection of antibodes against SARS-CoV using BHK-21 cells stably expressing recombinant viral nucleoprotein

Objective: To develop an indirect immunofluorescence assay for detection of antibodies against of SARS-CoV in ordinary laboratory by using BHK-21 cells stably expressing SARS-CoV nucleoprotein.Methods:Recombinant plasmid containing SARS-CoV nucleoprotein (N)gene was transfected into BHK cells to establish the BHK cell lines stably expressing the N protein. Specificity of the immunofluorescence method using the engineered cell lines was validated with antibodies against SARS-CoV and other respirovirus. Results and Conclusion: The indirect immunofluorescence assay based on engineered BHK cells was developed and could detect specially the antibody against SARS-CoV.The method provides a rapid and safe means for dectection of SARS-CoV in ordinary laboratory.