| 流式细胞术快速检测杆状病毒滴度 |
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| 引用本文： | 徐鹏,张佑红,杨益,彭继明,陈龙,靖志强,危威,马静,秦琴.流式细胞术快速检测杆状病毒滴度[J].武汉化工学院学报,2010(1):57-60. |
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| 作者姓名： | 徐鹏 张佑红 杨益 彭继明 陈龙 靖志强 危威 马静 秦琴 |
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| 作者单位： | 武汉工程大学化工与制药学院; |
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| 基金项目： | 国家自然科学基金(20876120)资助项目 |
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| 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．
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| 关 键 词： | 流式细胞术 杆状病毒 SYBR Green I |
Rapid titration of baculovirus by flow cytometry |
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| XU Peng,YANG Yi,PENG Ji-ming,CHEN Long,JING Zhi-qiang,WEI Wei,MA Jing,QIN Qin,ZHANG You-hong.Rapid titration of baculovirus by flow cytometry[J].Journal of Wuhan Institute of Chemical Technology,2010(1):57-60. |
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| Authors: | XU Peng YANG Yi PENG Ji-ming CHEN Long JING Zhi-qiang WEI Wei MA Jing QIN Qin ZHANG You-hong |
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| Affiliation: | XU Peng,YANG Yi,PENG Ji-ming,CHEN Long,JING Zhi-qiang,WEI Wei,MA Jing,QIN Qin,ZHANG You-hong(School of Chemical Engineering , Pharmacy,Wuhan Institute of Technology,Wuhan 430074,China) |
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| Abstract: | 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. |
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| Keywords: | flow cytometry baculovirus SYBR Green I |
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