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本研究建立了流式细胞仪快速检测鲤春病毒血症病毒(spring viraemia of carp virus,SVCV)滴度的方法。运用荧光激活细胞分选(fluorescence-activated cell sorting,FACS)技术检测SVCV A1株对草鱼性腺细胞系(GCO)的感染情况。用SVCV病毒单克隆抗体为一抗,FITC标记的羊抗鼠抗体为二抗,运用FACS来检测感染后不同时间点,以及不同病毒接种量的阳性细胞率。感染第3天时为最佳的病毒滴度测定时间点,测得SVCV的病毒滴度为8.31×105 FIU/m L,最低检测病毒滴度为(1000 FIU/m L),与传统空斑试验(plaque assay,PA)相比,两种方法测得的结果基本一致。实验结果表明,FACS是一种简捷、高效、直接的检测SVCV滴度的方法,是一种新型的病毒滴度测定方法。
In this study, we established a rapid flow cytometry (FCM) method for detecting the titer of the virus of carp virus (SVCV). Fluorescence-activated cell sorting (FACS) was used to detect the infection of SVCV A1 strain in GCO. Using SVCV monoclonal antibody as the primary antibody, FITC-labeled goat anti-mouse antibody as the secondary antibody, using FACS to detect the different time points after infection, and the positive rate of virus inoculum. The optimal titer of virus at the third day of infection was 8.31 × 105 FIU / mL and the lowest virus titer (1000 FIU / mL) (plaque assay, PA) compared to the two methods measured results are basically the same. The experimental results show that FACS is a simple, efficient and direct method to detect SVCV titer, which is a new method for determination of virus titer.