目的 验证Taqman荧光定量PCR检测生物制品生产用细胞中逆转录病毒的方法。方法 从专属性、检测限、线性、重现性、耐用性、准确性、精密度几个方面验证该方法在质量控制中的可行性。结果 该方法可检测2BS细胞、Vero细胞中的逆转录病毒，检测限可达1.0×10³ pU/μl逆转录酶。该法线性良好，决定系数＞0.960。该方法重现性好。试剂经5次冻融，体系在室温放置1 h均不影响实验结果。外加标准品的回收率均高于70%，相对标准偏差＜5%。结论 Taqman荧光定量PCR法可用于检测用于2BS细胞和Vero细胞是否被逆转录病毒污染。

Objective  To validate Taqman quantitative PCR assay for the detection of retroviruses in cells for biological production. Methods  The feasibility of this method in quality control was verified in specificity, detection limit, linearity, reproducibility, robustness, accuracy and precision. Results  The method detected reverse transcriptase, reflecting retrovirus in human diploid cell 2BS strain and Vero cell. The detection limit was 1.0×10³ pU/μl reverse transcriptase. The method had good linearity with coefficient of determination >0.960. The method was reproducible. The experimental result was unaffected after the reagent was frozen and thawed for up to 5 times, and after the reaction mix was left at room temperature for 1 h. The recovery rate of added standard was >70%, and the relative standard deviation was <5%. Conclusions  Taqman real-time PCR method can be used to detect retrovirus in both human diploid cell 2BS strain and Vero cell.