检测流行性出血热病毒滴度和中和抗体效价的半微量空斑法

建立了检测流行性出血热病毒滴度和中和抗体效价的半微量空斑法。小牛血清与胎牛血清的培养效果无差异。7株不同来源的出血热病毒均能在E6细胞上形成空斑。接种的病毒浓度与形成的空斑数呈直线关系。用空斑法测得的病毒滴度稍低于荧光TCIE50滴定法。空斑减少中和试验的敏感性较荧光中和试验高30倍左右。同时还初步表明了本方法可用于流行性出血热病毒的抗原性分析。

A SEMIMICRO PLAQUE ASSAY FOR MEASURING VIRUS TITER AND DETECTING SPECIFIC NEUTRALIZING ANTIBODY OF EPIDEMIC HEMORRHAGIC FEVER

A semimicro plaque titration assay and plaque reduction neutralization test for epidemic hemorrhagic fever virus (EHFV) (hemorrhagic fever with renai syndrom virus ) has been established. All soven EHFV strains tested could form plaques on Vero-E6 cell line culture. An approximately linear relationship existed between the number of plaques and virus con-centration in the inoculum, showing the reliability of this assay. The virus titer measured by the plaque assay was a little lower than that measured by immuno-fluorescent TCID50 assay, probably due to less efficient virus absorption in the plaque assay. The neutralizing antibody tested by PRNT has been shown to be approximately thirty two-fold higher than that of immuno-fluorescent neutralizing test. Preliminary result also showed that by using this semimicro PRNT method, antigenic differences among EHF vivus strains could be demonstrated.