Colti病毒抗原制备、保存及其在检测患者血清Colti病毒抗体中的应用  

Preparation and storage of coltivirus antigen and its application in detection of Coltivirus antibodies in serum from patients

作　　者：陶三菊[1] 王焕琴[1] 曹玉玺[1] 杨冬荣[1] 刘琴芝[1] 徐丽宏[1] 何英[1] 陈伯权[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《中华实验和临床病毒学杂志》2002年第3期232-235,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的　获得较高滴度和稳定的Colti病毒抗原 ,进一步开展Colti病毒感染的血清学诊断。方法　用C6 36细胞培养Colti病毒 ,于不同时间收获后进行病毒滴度测定。病毒经聚乙二醇(PEG)纯化浓缩后抗原分别保存于 - 2 0℃和 4℃备用。利用该抗原采用ELISA法检测患者血清标本的Colti病毒抗体。结果　在制备Colti病毒抗原时 ,以细胞培养 3～ 4周的病毒滴度最高 ,经PEG纯化浓缩的抗原于 - 2 0℃和 4℃条件下 ,保存 6个月 ,其抗原滴度仍保持在较高水平。特别是加入甘油的抗原无论在 - 30℃ ,还是在 4℃下 ,甚至可保存 2年。用这些抗原 ,检测疑似乙型脑炎或病毒性脑炎患者标本 114 1份 ,Colti病毒IgM抗体阳性 130例 ,阳性率为 11.4 % (130 114 1) ,特别是检测广州市儿童医院患者标本 4 1份 ,9份Colti病毒抗体阳性 ,阳性率为 2 2 .0 % ,其中 5例临床诊断为病毒性脑炎。结论　为建立检测Colti病毒抗体及诊断试剂盒提供了较稳定的抗原 ,Colti病毒可能是引起我国夏、秋季脑炎的又一重要病原。Objective To prepare purified and concentrated coltivirus high titer antigen in order to further detect antibodies against coltivirus in serum sample of patients. Methods The coltivirus in C6/36 cells was cultured and harvested at different time,and the titer was titrated.The virus was purified and concentrated by polyethylene glycol (PEG), and stored at -20℃ and 4℃, with and without glycerol, respectively, then the titer of coltivirus antigen was tested by indirect ELISA. By using the antigen, coltivirus antibodyies in serum samples from both suspected Japanese encephalitis (JE) and viral encephalitis(VE) patients were detected. Results The highest titer of coltivirus was found at 3 4 weeks of culturing. The antigen titer cauld be maintained at least for 6 months, especially antigen with glycerol either at 4℃ or at -30℃ even for two years.To tally 1 141 serum samples from patients diagnosed clinically as JE and VE were tested. The results showed that 130 samples were coltivirus IgM antibody positive and the average positive rate was 11.4%(130/1 141). Among 41 samples of paired serum from patients in Guangzhou Children's Hospital,9 samples were positive,the positive rate was 22.0%(9/41) in which 5 samples were diagnosed clinically as VE. Conclusion Stable and purified coltivirus antigen was obtained in order to test coltivirus antibodies as well as development of kits. Coltivirus probably can cause summer autumn encephalitis in China.

关 键 词：COLTI病毒 抗原 制备 保存 血清学诊断 

分 类 号：R446.61[医药卫生—诊断学]