提纯CVB1的方法学改进和CVB1的灭活  

Improved Method for CVB1 Purification and Its Inactivation

作　　者：陈国俊[1] 刘焯霖[1] 张成[1] 陈元[1] 方丹云[1] 吴金浪[1] 

机构地区：[1]中山医科大学,广州510080

出　　处：《中山大学学报（医学科学版）》1999年第S1期4-7,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金;卫生部临床重点学科基金

摘　　要：目的：研究聚乙二醇沉淀和蔗糖梯度离心法提纯柯萨奇病毒Ｂ１（ＣＶＢ１） 的效率，并采用β丙内酯（ＢＰＬ） 将其灭活，观察ＢＰＬ的灭活效果。方法：①聚乙二醇沉淀法将病毒悬液初步浓缩；②在１０％ ～５０％ （ ｍ ／ Ｖ） 蔗糖梯度中上样，低温超速离心；③电镜观察并做病毒活性测定；④ＢＰＬ对病毒进行灭活，测定残存毒力。结果：①病毒在５０％ （ ｍ ／ Ｖ） 蔗糖处有一吸收峰，并被电镜证实；②ＢＰＬ灭活病毒彻底，灭活后病毒稀释１０２ ～１０１０ 倍接种细胞未见病变。结论：ＣＶＢ１ 经聚乙二醇沉淀和蔗糖梯度超速离心后收获率高，ＢＰＬ灭活彻底，保证了进一步实验的成功。Objective: To study an improved method of Coxsackie virus B1(CVB1) purification with polyethylene glycol (PEG) sedimentaion followed by sucrose ultracentrifugation ,and to study the effect of virus inactivation by β propiolactone (BPL). Methods: ① Virus suspension was concentrated primarily by PEG.② Samples were layered onto 10%～50% sucrose gradient and purified by ultracentrifugation. ③ Virues were identified under electronic microscope, and the toxicity was measured as well. ④ Viruses were inactivated by BPL and the toxicity measured again. Results: ① CVB1 showed a peak value of A 260  in 50% sucrose gradient, which was approved by electronic microscope. ② CVB1 were thoroughly inactivated by β propiolactone, cytopathology was not seen even when viruses were diluted from 10 2 to 10 10 times. Conclusions: ① Highly purified CVB1 after PEG sedimentation and sucrose ultracentrifugation was obtained. ② Inactivation of CVB1 by BPL was complete, which guarantee the successfulness for further studies.

关 键 词：柯萨奇病毒B组/分离和提纯 丙内酯 离心法 梯密度 病毒灭活 

分 类 号：R373.23[医药卫生—病原生物学]