肠病毒71型空斑检测方法的建立  

Development of Plaque Assay for Enterovirus 71

作　　者：郝春生[1] 赵敏[1] 张晨[1] 何薇薇[1] 王潇潇[1] 杨永娟[1] 张中洋[1] 沈心亮[1] 李秀玲[1] 

机构地区：[1]北京生物制品研究所,北京100024

出　　处：《中国生物制品学杂志》2009年第10期1023-1025,共3页Chinese Journal of Biologicals

基　　金：国家科技支撑计划(2008BAI69D00)

摘　　要：目的建立肠病毒71型(EV71)空斑检测方法,为EV71生物学特性及疫苗的研究提供手段。方法将不同稀释度的5株EV71分别接种至单层RD细胞和Vero细胞,加入甲基纤维素覆盖,计数空斑,并计算病毒滴度。通过3次连续检测,验证空斑法的精密性,并对空斑法与微量细胞病变法检测病毒滴度的结果进行比较。结果采用RD细胞和Vero细胞,通过空斑法对EV71病毒滴度进行检测,96h后均能规律地出现边缘整齐、清晰的空斑,RD细胞和Vero细胞检测的空斑直径分别为1～2mm和0.5～1mm。连续3次重复试验结果显示,试验间变异系数的平均值Vero细胞为2.52%,RD细胞为4.49%。空斑法与微量细胞病变法比较,其检测结果具有良好的线性相关性,相关系数为r=0.972,P=0.006。结论已建立了精密性好、出斑规律的EV71病毒空斑检测方法。Objective To develop a plaque assay for enterovirus 71（EV71）and provide a method for study on biological property of EV71 and preparation of EV71 vaccine.Methods The EV71 at various concentrations were inoculated onto RD and Vero cell monolayers respectively,then added with methylcellulose and counted for plaque,based on which the virus titer was calculated.The precision of developed method was verified by 3 tests,and the determination result of virus titer by the method was compared with that by micro-CPE method.Results Clear plaques with regular borders were observed in both RD and Vero cells 96 h after inoculation with EV71,at sizes of 1 ～ 2 and 0.5 ～ 1 mm respectively.The variation coefficients of results of 3 tests on Vero and RD cells were 2.52% and 4.49% respectively.The determination result by the developed plaque assay showed good linear relationship to that by micro-CPE（r = 0.972,P = 0.006）.Conclusion A precise and regular plaque assay for EV71 was developed.

关 键 词：肠病毒71型 空斑试验 病毒滴定 

分 类 号：Q-33[生物学]