朱明华 戴益民 200433 上海第二军医大学病理学教研室 中华实验和临床病毒学杂志 1999 0 0 1


    关键词：载体，逆转录病毒；基因治疗；支原体 期刊 zhsyhlcbdxzz 0 论著 fur -->


    

【摘要】 目的 在以逆转录病毒为载体的抗乙型肝炎病毒(HBV)细胞内免疫基因治疗研究过程中，建立较完整的生物安全检测系统，为临床应用奠定基础。方法 包括无菌，支原体和可复制性逆转录病毒的检测。支原体检测采用聚合酶链反应(PCR)方法，可复制性逆转录病毒的检测应用S⁺ /L^- 试验，NIH3T3细胞扩增试验和neo基因补救分析。结果 在所建立的靶基因包装细胞系中，无菌试验均为阴性，一株包装细胞支原体呈阳性，全部包装细胞系未测到可复制性逆转录病毒。结论 本研究采用的生物安全检测系统具有稳定性好，敏感染性高的优点，对基因治疗临床试验的生物安全检测具有重要的应用价值。

    Bio-safety testing for retroviral vector as gene therapy delivery system ZHU Minghua, DAI Yimin. Department of Pathology, Second Military Medical University, Shanghai 200433

    【Abstract】 Objective To generate a bio-safety testing system including the testing of sterility, mycoplasma and replication-competent retroviruses(RCR) in the study of gene therapy anti-hepatitis B virus by intracellular immunization.Methods Mycoplasma was detected by polymerase chain reaction. S⁺ /L^- assay, NIH3T3 amplification and rescue assay of neo gene were performed to determine RCR. Results The results showed that all of the packaging cell lines were negative for aerobic, anaerobic bacteria and fungi. One of the packaging cell lines was positive for mycoplasma. No RCR was detected in all of the packaging cell lines.Conclusions The results suggested that the methods using in present report are stable and sensitive, it is very useful for bio-safety testing of gene therapy in clinical trials.

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