重组人1型单纯疱疹病毒样品的活性检测研究

目的： 建立分析重组人1型单纯疱疹病毒（rHSV-1）样品感染滴度和生物学活性的方法。方法： 采用噬斑法检测重组人1型单纯疱疹病毒的感染滴度，采用选择性增殖活性检测法和肿瘤细胞杀伤活性检测法检测样品的生物学活性，分别对上述方法进行重复性和精密度性能验证，并用上述方法对6批样品中的感染滴度和生物学活性进行检测。结果： 经验证，上述3种方法均有较好的重复性和精密度，精密度RSD分别为5.3%、3.9%和19.4%；6批测试样品中，感染滴度为（3.06±0.37）×10⁷ PFU·mL^-1，选择性增殖活性为（1.58±0.63）×10⁵，肿瘤细胞杀伤活性为0.027 8±0.015 9。结论： 经方法学验证，噬斑法、选择性增殖活性检测法和肿瘤细胞杀伤活性检测法可分别用于重组人1型单纯疱疹病毒的感染滴度和生物学活性的检测。

Objective: To develop methods to determine the biological activity and infectious titer of recombinant human herpes simplex virus type 1(rHSV-1) samples. Methods: The titer of rHSV-1 was determined by plaque assay, and the biological activity of rHSV-1 was determined by selective proliferative assay and tumoricidal assay. All the methods were validated in repeatability and precision, and used to detect the titers and biological activities of 6 batches of rHSV-1 samples. Results: The developed methods showed good repeatability and precision, with inter-assay precision being 5.3%, 3.9% and 19.4%, separately. Six batches of rHSV-1 were determined, with the average titer being (3.06±0.37)×10⁷ PFU·mL^-1, the average selective proliferative activity being (1.58±0.63)×10⁵, and the average tumoricidal activity being 0.027 8±0.015 9. Conclusion: The developed methods are validated to be viable for the determination of titer and biological activity of rHSV-1.

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