目的：通过对比两种菌种保存方法的验证结果，选择出一种保存时间长、占用空间少、操作安全、出错风险低、菌种稳定性好的方法，为无菌药品生产企业微生物实验室菌种保存提供参考。方法：使用金黄色葡萄球菌、铜绿假单胞菌、生孢梭菌、枯草芽孢杆菌、白色念珠菌、黑曲霉这6种标准菌种和1种从洁净环境分离的藤黄微球菌，通过复苏率、形态学、染色学和数值分类微生物鉴定法，分别对液体石蜡覆盖保存法和10%甘油-生理盐水冷冻管保存法进行验证。结果：经菌种保存的方法学验证，液体石蜡覆盖保存法菌种保存时限最短仅为6个月，而使用10%甘油-生理盐水冷冻管保存法，细菌和酵母菌能保存24个月，霉菌保存18个月。结论：10%甘油-生理盐水冷冻管保存法优于液体石蜡覆盖保存法，可用于微生物实验室的菌种保存。

Objective:By comparing the validation results of two strain preservation methods, a persistent, space-saving, safe, error-avoiding, and stable method was found for strain preservation so as to provide references for microbial laboratories of aseptic pharmaceutical manufacturers. Methods:The liquid paraffincovered preservation method and 10% glycerol-saline preservation method were verified by testing recovery rate, morphology, staining and numerical classification mcrobiological identification system using 6 standard strains (Staphylococcus aureus, Pseudomonas aeruginosa, Clostridium sporogenes, Bacillus subtilis, Candida albicans and Aspergillus niger) and Micrococcus luteus isolated from clean environment. Results:The strains could only be stored for 6 months using the liquid paraffin-covered preservation method. Whereas, the bacteria and yeast could be stored for 24 months, and the mould for 18 months using 10% glycerol-saline method. Conclusion:10% glycerol-saline conservation method was better than the liquid paraffin-covered preservation method and could be used for strain preservation in microbial laboratories.