逆转录病毒介导的基因治疗中的安全性检测

逆转录病毒介导的基因治疗中安全性的最大危险是产生有复制能力的逆转录病毒（RCR）。本实验对经3T3扩增的样品进行 S~+L~-分析、标记拯救分析和PCR扩增方法检测RCR。比较三种方法,标记拯救分析比S~+L~-分析结果容易判断,且PCR可以从10~5个无病毒基因的细胞中检测出一个带有病毒基因的细胞。3T3扩增可提高灵敏度约10倍。安全性是基因治疗首要考虑的问题,本方法为其提供了保证。

Safety Test in Retroviral-mediated Gene Therapy

Replication of competent retrovirus(RCR) is the primary danger in retro-viral-mediated gene transfer involved gene therapy. In this study, we used S+L- assay, marker rescue assay and PCR to detect RCR after 3T3 amplification. Comparing the three methods, We found that the marker rescue assay was easy-manipulated and PCR was sensitive capable of detecting one marked cell among 105 unmarked cells. Since safety must be first considered before clinical trials of human gene therapy, the three assays could be useful for monitoring against RCR in retroviral-mediated gene therapy.