荧光定量聚合酶链反应检测移植后血细胞嵌合率的体外实验研究

Real-time fluorescence quantitative polymerase chain reaction to detect chimerism rate after allotransplantation

目的 建立一种定量检测移植后血细胞嵌合率的方法 ,以监测吻合血管同种异体骨移植受体的免疫耐受状态。方法 结合动物实验模型 ,设计性别决定因子Y(SRY)实时荧光定量聚合酶链反应 (PCR)检测血细胞嵌合率的方法 ,建立质控标准品 ,使用实时荧光定量PCR技术 ,获得标准曲线。结果 标准曲线为一条严格的直线型曲线 ,线性相关系数r >0 .98,反映了标准品的数量关系 ,敏感度可达 0 .3 % ,符合技术要求。结论 本实验方法标准可靠 ,可用来检测实验样品的血细胞嵌合率。

Objective To establish a method to detect chimerism rate in peripheral blood after transplantion to monitor the state of immunological tolerance of vascularized bone allotransplantation recipients.Methods A model of animal experiment,a kit for sexdeter mining region Y (SRY) and a set of standard sample were developed for drawing standard curve by real time fluorescence quantitative PCR.Results Standard curve was a strict beeline with correlation coefficient above 0.98.Conclusion This method is accurate and reliable,and can be used to detect chimerism rate of the experimental samples.