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目的:建立用流式细胞仪快速测定汉滩病毒滴度的方法。方法:汉滩病毒76-118株感染Vero-E6细胞,以汉滩病毒核蛋白单克隆抗体(mAb)3G1为一抗,FITC标记的羊抗小鼠抗体为二抗,用流式细胞术(FCM)检测阳性细胞率,评价感染后不同时间点,以及不同病毒接种量的阳性细胞率,并与间接免疫荧光法对比。结果:感染后36 h阳性细胞百分率为(10.06±0.42)%,最低检测的病毒滴度为100 TCID50/mL。结论:相对于传统的空斑实验及间接免疫荧光滴定法,FCM是一种简单、快速、有效检测汉滩病毒滴度的方法。
Objective: To establish a method for rapid determination of Hantaan virus titer by flow cytometry. Methods: Hantaan virus strain 76-118 was infected with Vero-E6 cells. The primary antibody of Hantaan virus nucleoprotein monoclonal antibody (mAb) 3G1 was screened. FITC-labeled goat anti-mouse antibody was used as secondary antibody. FCM) were used to detect the positive cell rate, evaluate the different time points after infection, and the positive rate of different virus inoculation quantity, and compared with indirect immunofluorescence method. Results: The percentage of positive cells at 36 h after infection was (10.06 ± 0.42)%, and the lowest titer of virus was 100 TCID50 / mL. Conclusion: Compared with traditional plaque assay and indirect immunofluorescence titration, FCM is a simple, rapid and effective method for detecting Hantaan virus titer.