目的 建立中国仓鼠卵巢（Chinese hamster ovary，CHO）细胞收获液的小鼠微小病毒（minute virus of mice， MVM）荧光定量PCR（quantitative PCR，qPCR）检测法并进行方法学验证及初步应用。方法 采用qPCR法对CHO细胞培养收获液进行MVM检测，验证该方法的重复性、中间精密度、专属性、耐用性和检测限。对12份CHO细胞收获液进行MVM检测。结果 同一样品重复检测 6次，加标样品循环阈值的变异系数为0.6%~0.8%。不同人员间的变异系数为0.7%。与猪细小和猪圆环病毒无交叉反应。当细胞数在1.0×10⁷、1.1×10⁷、9.0×10⁶波动时检测结果未受影响。检测限为200 拷贝/μL。采用该方法对12份CHO细胞收获液进行检测，均未检出MVM。结论 建立的qPCR法具有良好的重复性、中间精密度、专属性、耐用性和检测限，可以用于CHO细胞收获液中MVM的检测。

Objective To establish and verify the quantitative PCR (qPCR) detection method for mouse minute virus (MVM) in Chinese hamster ovary (CHO) cell harvest, and carry out preliminary application.Methods CHO cell culture harvest was tested for MVM by qPCR, and the repeatability, intermediate precision, specificity, robustness and detection limit of the method were verified. The method was used to test 12 CHO cell harvests for MVM.Results The coefficients of variation (CVs) of cycle threshold of the spiked samples tested 6 times were 0.6%-0.8%, and the CV of different operators was 0.7%. There was no cross-reactivity with porcine minute virus or porcine circovirus. The results were not affected when cell number varied at 1.0×10⁷, 1.1×10⁷, and 9.0×10⁶. The detection limit was 200 copies/μL. No MVM was detected in 12 CHO cell harvests by this method.Conclusion The established qPCR method has good repeatability, intermediate precision, specificity, robustness and detection limit, and can be used for the detection of MVM in CHO cell harvest fluid.