Abstract: Objective To compare the detection effects of culture method, Gram stain microscopy and 16S rRNA PCR on bacterial detection in fecal samples of germ-free mice,provide a basis for the selection of germile murine bacteria detection method. Method The culture method, Gram stain microscopy and 16S rRNA PCR method were used to detect the feces of GF mice, the feces of SPF mice and the test samples with known bacterial content ( the feces of germ-free mice were mixed with Staphylococcus aureus and Escherichia coli, artificially made) , and the detection limits of the three method on bacteria in the feces of germ-free mice were verified. Three method were used to detect clinical samples at the same time, the advantages and disadvantages of the three detection method were compared, and the feasibility of the three method in actual detection was analyzed. Result The sensitivity of culture method, Gram stain microscopy and 16S rRNA PCR method was 10²CFU / g, 10⁸CFU / g and 10⁶CFU / g, respectively, and the positive rate of clinical samples was 1. 36% ( 6 / 441 ) . Conclusion Culture method, Gram staining microscopy and 16S rRNA PCR method have their own advantages and disadvantages, and they need to be used in combination to learn from each other ’ s strengths to ensure the reliability of the detection result.

Key words: germ-free mice, sterility test, cultivation method, PCR, gram staining microscopic examination

摘要： 目的 比较培养法、革兰染色镜检法和 16S rRNA PCR 方法对无菌小鼠粪便样品细菌检测的检测性能,为无菌鼠细菌检测方法的选择提供依据。 方法 利用培养法、革兰染色镜检法、16S rRNA PCR 方法,分别检测 GF 小鼠粪便、SPF 小鼠粪便以及已知含菌量的测试样品(无菌小鼠粪便与金黄色葡萄球菌、大肠埃希菌混合,人为制作而成) ,验证 3 种方法对无菌小鼠粪便中细菌的检测限度;并同时使用 3 种方法检测临床样品,对比 3 种检测方法的优缺点,分析 3 种方法在实际检测中的可行性。 结果 培养法、革兰染色镜检法和 16S rRNA PCR 方法检测灵敏度分别为 10²CFU / g、10⁸CFU / g 和 10⁶CFU / g,临床样品检测中阳性率均为 1. 36%(6 / 441) 。 结论 培养法、革兰染色镜检法、16S rRNA PCR 法 3 种检测方法各有优缺点,需要联合使用,取长补短,确保检测结果的可靠。

关键词: 无菌小鼠, 细菌检测, 培养法, PCR, 革兰染色镜检