【摘要】 目的探讨丙型肝炎病毒样颗粒(HCVLPs)在亚甲蓝光化学灭活处理过程中的变化趋势及其作为亚甲蓝光化学法灭活HCV效果评价物的可行性。方法以HCV阳性血浆作为平行对照组(HCV RNA载量约6.53 logcopies/m L)(n=6),将HCVLPs用代血浆调整成合适浓度(HCV RNA定量约为6.74 logcopies/m L)的悬液作为实验组(n=5),将2组分装至PVC血袋中,MB+L 1μmol/L进行病毒灭活处理,分别于光照处理0、5、10、20、30 min时取样,用荧光定量PCR技术检测病毒核酸载量;同时以细胞病变法测定HCV模型病毒sindbis病毒的残余滴度,验证MB+L灭活HCV的效果。结果模型病毒sindbis的病毒滴度降至检测限以下(log TCID50/m L≤0.5);MB+L处理过程中,随着光照处理时间的增加,HCV及HCVLPs的核酸载量明显下降,分别从6.53与6.74下降至4.51和2.89log copies/m L(P<0.05),且2组在不同取样点的HCV RNA载量之间具有相关性(R2>0.98,Significance F<0.01)。结论 HCVLPs能够反映MB+L灭活处理中HCV RNA动态变化,或有望成为安全而有效的HCV灭活评价物来监控HCV灭活效果。更多还原

【Abstract】 Objectives To investigate the dynamic changes in HCV virus-like particles during photochemical inactivation of MB + L,and to explore the feasibility of HCV virus-like particles(VLPs) on evaluating the effectiveness of HCV inactivation.Methods HCV VLPs were adjusted to proper concentration(at a viral load of 6.74 log copies / m L),with HCV positive plasma as control(at a viral load of 6.53 log copies / m L).Both VLPs suspension and plasma were separated into PVC bags and treated with MB + L.Viral loads of VLPs and HCV were tested for 0,5,10 or 30 minutes of light treatment by Real-Time PCR.The residual infectivities of HCV virus and Sindbis virus were determined by evaluating their cytopathic effect.Results The infectivity of Sindbis virus was determined to be below the detection limit(log TCID50/ m L≤0.5),indicating that HCV was effectively inactivated.Meanwhile,viral loads of VLPs and HCV decreased significantly(P < 0.05) as the time of MB + L treatment lengthened,from 6.53 and 6.74 to4.51 and 2.89 log copies / m L,respectively.The decline of viral load of VLPs and HCV showed a significant correlation(R2>0.98,Significance F < 0.01).Conclusion The results indicated that HCV VLPs could reflect the effect on HCV inactivation by methylene blue with visible light and might be safe and effective for evaluating the effects on virus inactivation.更多还原