SARS-CoV的培养和灭活条件的研究  

Conditions for SARS-CoV cultivation and inactivation

作　　者：张松乐[1] 马丽娟[1] 田光[1] 张良艳[1] 张雪颖[1] 王希良[1] 

机构地区：[1]军事医学科学院微生物流行病研究所免疫室,北京100071

出　　处：《中华实验和临床病毒学杂志》2005年第2期135-137,i001,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家"8 63"基金资助项目 (2 0 0 3AA2 0 82 2 4) ;军队"十五"重大攻关课题资助项目 (0 3F0 0 3 3 )

摘　　要：目的　研究SARS CoV的大量培养和灭活方法,为制备马抗SARS免疫球蛋白制剂提供安全的病毒。方法　使用SARS CoV感染Vero、Vero E6和2BS细胞株,筛选对SARS CoV较敏感的细胞株和最佳病毒感染量;在2 5℃、33℃和37℃培养条件下,检测SARS CoV的最适培养温度。在室温下,使用1∶2 0 0 0～1∶2 0 0 0 0的β丙内酯浓度灭活SARS CoV ,观察最佳灭活时间和效果。结果　SARS CoV对Vero和Vero E6细胞株比较敏感。在37℃、5 %CO2 孵箱中培养72h ,细胞病变达75 %以上。1∶4 0 0 0的β丙内酯能够完全灭活SARS CoV。将灭活后的病毒液在2℃～8℃作用16h ,37℃水浴2h能够完全使β丙内酯的毒性水解。结论　用Vero或Vero E6细胞培养SARS CoV ,在37℃、5 %CO2 孵箱中培养72h ,病毒滴度最高。用1∶4 0 0 0的β丙内酯浓度加入病毒液中,作用1h能够完全灭活SARS CoV。Objective To study the method for cultivation and inactivation of SARS-CoV. Methods In order to choose the sensitive cell strain and the best infection dose of the virus, Vero, Vero-E6 and 2BS cell lines were infected with SARS-CoV. The cultivation temperature was selected among 25℃,33℃ and 37℃. The best inactivating time and effect were observed with β-propiolactone whose concentration ranged from 1∶2 000 to 1∶ 20 000 at room temperature. Results Vero and Vero-E6 cell lines were sensitive to SARS-CoV. The cytopathic changes of the cells were 75% at 37℃ in 5% CO 2 incubator after infection. SARS-CoV was inactivated completely in β-propiolactone(1∶4 000). The toxicity of β-propiolactone was hydrolyzed completely when the inactivated virus was cultured for 16 hours at 2℃～8℃and in water bath for 2 hours at 37℃. Conclusion The titer of SARS-CoV was the highest when it was cultured in Vero or Vero-E6 cells for 72 hours at 37℃ in 5% CO 2 incubator. SARS-CoV was inactivated completely in β-propiolactone when its concentration was 1∶4 000 and the interaction time was 1 hour.

关 键 词：养和 SARS-COV感染 Vero-E6 β-丙内酯 免疫球蛋白制剂 SAILS 37℃水浴 细胞株 灭活方法 培养条件 培养温度 灭活时间 细胞病变 细胞培养 病毒滴度 72h CO2 病毒液 2BS 感染量 最佳 敏感 浓度 

分 类 号：TQ464[化学工程—制药化工]