检测轮状病毒滴度的荧光灶法的改进  

Improvement of Fluorescent Focus Assay for Quantitation of Rotavirus

作　　者：梁婧[1] 徐葛林[1] 黄晓媛[1] 王继麟[1] 孙文[1] 夏飞[1] Kapikian AZ 杨晓明[1] 

机构地区：[1]武汉生物制品研究所,武汉430060 [2]Epidemiology Section,Laboratory of Infectious Diseases,National Institute of Allergy and Infectious Diseases,National Institutes of Health

出　　处：《中国生物制品学杂志》2008年第7期620-622,共3页Chinese Journal of Biologicals

基　　金：十一五863计划生物和医药技术领域疫苗与抗体工程重大项目(2006AA02A210)

摘　　要：目的对检测轮状病毒滴度的荧光灶试验(FFA)方法进行改进。方法以FFA法检测轮状病毒滴度,并观察残留血清、病毒吸附量、甲基纤维素、离心以及Ca2+等因素对检测结果的影响,并验证改进后FFA的重复性。结果残留血清、病毒接种量、吸附后不洗涤、PBST洗板对FFA检测结果均无影响。而接种病毒后离心及不加甲基纤维素覆盖与对照组比较,差异有显著意义。用改进后方法对同批病毒进行18次试验,CV值为1.7%。结论改进后的方法具有操作简便、结果稳定、重复性好等优点。Objective To improve the fluorescent focus assay （FFA） for quantitation of rotavirus. Methods Determine the titer of rotavirus strain MRV1 by FFA, and observe the influence of residual serum, virus adsorption quantity, methyl cellulose, cen- trifugation and calcium ion on the determination result. The improved FFA were evaluated for reproducibility. Results Residual serum, virus loading quantity, deletion of washing step after adsorption, washing plate with PBST showed no significant influence on the determination result. However, centrifugation after virus inoculation and adding no methyl cellulose showed significant influence on determination resuh. The titers of virus samples of the same batch were determined by the improved FFA for I8 times, and the CV value of results was 1.7%. Conclusion The improved FFA was simple, stable and of good reproducibility.

关 键 词：轮状病毒 荧光灶试验 定量 滴度 

分 类 号：R392-33[医药卫生—免疫学] R373[医药卫生—基础医学]