应用Real Time PCR方法检测病毒灭活去除效率

Real time PCR as a method to cletect virus inactivation and clearance efficiency

目的建立检测血液制品中病毒灭活去除效率的模型。方法应用SYBR GREEN I实时荧光定量PCR验证不同方法的病毒去除效率,并用病毒感染力实验验证不同方法的病毒灭活效率。结果以PCR产物为外参的实时荧光定量PCR正确反映出实际核酸含量和检出拷贝数的线性关系,以重组质粒为外参的实时荧光定量PCR实验证明巴氏灭毒法的病毒去除效率低于金磁颗粒法。病毒感染力实验证明巴氏灭毒法灭活效率略高于金磁颗粒法。结论实时荧光定量PCR法联合病毒感染力试验可以有效评价不同方法在病毒灭活和病毒去除方面的差异。

Aim To evaluate virus clearance efficiency in plasma product.Methods Using SYBR green I dye a Real Time PCR is applied to detect the virus quantity variety between samples which have been treated with different protocols.Inactivation efficiency is cvaluated by virus titer test.Results Detected copy numbers of virus is highly consistent with the exact amount of virus.Different protocols have highly different efficiency on virus removal.Conclusion It indicates the Real-time PCR technique can be engaged into the evaluation system of virus inactivation and removal.