流式细胞术快速检测杆状病毒滴度  

Rapid titration of baculovirus by flow cytometry

作　　者：徐鹏[1] 张佑红[1] 杨益[1] 彭继明[1] 陈龙[1] 靖志强[1] 危威[1] 马静[1] 秦琴[1] 

机构地区：[1]武汉工程大学化工与制药学院,湖北武汉430074

出　　处：《武汉工程大学学报》2010年第1期57-60,共4页Journal of Wuhan Institute of Technology

基　　金：国家自然科学基金(20876120)资助项目

摘　　要：为了快速而准确的测定杆状病毒的滴度，采用流式细胞术对经SYBR Green I染色后的杆状病毒直接计数．考察固定、破膜和染色等因素对检测结果的影响，并验证改进后的方法的重复性和线性性．杆状病毒最佳染色条件：质量分数为0．1％的多聚甲醛固定病毒30min后，加入一定量的SYBR Green I在80℃下孵育10min．测量结果CV值为2．4％（n=8），R^2为0．9998．整个测量过程由原来终点稀释法的7～10d缩短到1h．To develop a rapid and accurate method for baculovirus titration, flow cytometry （FCM） was adopted to quantitate baculovirus directly after staining with SYBR Green I. The effects of different treatment steps （fixation, permeabilization, staining, etc. ） were examined. The linearity and the reproducibility of the developed method were evaluated. The optimum dyeing conditions were shown： viral samples fixed with 0. 1% paraformaldehyde for 30 rain at 4 ℃, stained with SYBR Green I （1 × 10^-4 commercial concentration） for 10 rain at 80 ℃. The CV value of results is 2.4%（n=8）, R^2 value of different dilutions is 0. 999 8. The entire procedure is shortened to 1 h compared with the 7-10 d of end-point dilution method.

关 键 词：流式细胞术 杆状病毒 SYBR Green I 

分 类 号：Q331[生物学—遗传学]