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目的探讨麻腮风水痘联合减毒活疫苗(measles,mumps,rubella and varicella combined vaccine,live,MMRV)中水痘病毒在人二倍体细胞(MRC-5和2BS)上蚀斑的形成情况,建立不中和法检测MMRV中水痘病毒滴度,并进行验证。方法观察麻疹、腮腺炎和风疹病毒在人二倍体细胞上形成的细胞病变形态,以及风疹病毒对水痘病毒蚀斑形成的影响;通过完全中和、部分中和以及未中和等方法,检测MMRV中水痘病毒的滴度,最终建立不中和法检测水痘病毒滴度,并对方法进行重复性和准确性验证。结果风疹病毒不高于4.0 lg CCID50/孔时,水痘病毒在MRC-5或2BS细胞中产生的蚀斑形态发生微小改变,但蚀斑数量与完全中和风疹病毒后差异无统计学意义(P>0.5)。采用完全中和、部分中和以及未中和等方法检测MMRV中水痘病毒的滴度差异无统计学意义(P>0.5),且不中和法检测水痘疫苗病毒滴度的重复性(RSD为2.3%)和准确性(回收率为97.8%~100%)均较好。结论在人二倍体细胞(2BS或MRC-5)上,不使用抗血清中和直接进行四联疫苗中水痘病毒的滴度检测完全可行,可替代MMRV中经典的使用抗体中和的病毒滴定方法。
Objective To investigate the formation of plaque on human diploid cells (MRC-5 and 2BS) in measles, mumps, rubella and varicella combined vaccine, live, MMRV Non-neutralization assay of varicella virus in MMRV titers and verification. Methods The morphology of the cytopathic effect of measles, mumps and rubella virus on human diploid cells was observed and the effect of rubella virus on plaque formation of varicella was observed. By means of complete neutralization, partial neutralization and unneutralization, The titer of varicella virus in MMRV and the establishment of a non-neutralization assay for varicella virus titer were finally established. The repeatability and accuracy of the method were verified. Results When the rubella virus was no higher than 4.0 lg CCID50 / well, the plaque morphology of varicella virus was slightly changed in MRC-5 or 2BS cells, but there was no significant difference in the numbers of plaque after completely neutralizing rubella virus (P > 0.5). The titer of varicella virus in MMRV was not significantly different (P> 0.5) by using methods of complete neutralization, partial neutralization and non-neutralization, and the repeatability of varicella virus titer was not detected by neutralization assay (RSD 2.3%) and accuracy (recovery of 97.8% ~ 100%) are better. Conclusions The titer detection of varicella virus in the quadruple vaccine without the use of antiserum neutralization is completely feasible in human diploid cells (2BS or MRC-5) and can replace the classic use of antibody-neutralizing virus titration in MMRV method.