论文部分内容阅读
目的 研究GFP标记的复制缺陷型腺病毒在消毒剂灭活病毒试验中应用的可行性.方法 采用商品化Ad-easy系统经同源重组包装GFP标记的复制缺陷型腺病毒,扩增后进行荧光计数和复制缺陷性能检测,其作为指示病毒进行病毒灭活试验.结果 腺病毒基因同源重组后转染AD-293细胞出现绿色荧光和细胞病变,成功包装出病毒颗粒,扩增后经荧光计数病毒滴度为4.20×106 gfu/ml.盲传3代后,感染Vero细胞未出现荧光和细胞病变,表明无子代病毒的产生.以有效氯含量200 mg/L消毒剂作用于腺病毒20 min后,杀灭对数值为4.10.结论 荧光标记的复制缺陷型腺病毒在含氯消毒剂对病毒杀灭能力检测评价中具有一定的可行性,更为高效和安全.“,”Objective To investigate the disinfection effect of recombinant replication-defective adenovirus with fluorescent reporter in inactivation of virus.Methods The recombinant replication-defective adenovirus with fluorescent reporter was rescued by homologous recombination and transfection with commercialized Ad-easy system.The quality of recombinant virus was determined by fluorescence counting and replication-defection testing.The efficacy of recombinant adenovirus was evaluated by quantitative inactivation test.Results The recombinant adenovirus was successfully rescued with the presence of green fluorescent and cytopathogenic efficiency.The viral titer was 4.20 × 106 gfu/ml after amplification.No fluorescence and cytopathogenic efficiency were found after three generations of transfection,indicating no production of progeny virus.The average killing logarithm value of recombinant virus in suspension exposed to chlorine 200 mg/L for 20 min was 4.10.Conclusion The recombinant replication-defective adenovirus with fluorescent reporter can be feasible in the evaluation of virus inactivation test by shortening test period and improving safety.