菌种与培养基在抗生素微生物检定法中的应用分析

Investigation of Test Organisms and Media for Microbiological Assay of Antibiotics

目的考察菌种、菌种传代培养基及抗生素检定培养基对抗生素微生物检定法的影响,确定抗生素微生物检定法的系统适用性条件。方法选择美国菌种保藏中心标准菌株ATCC6633和中检院标准菌株CMCC(B)63501,分别用A、B企业的营养琼脂培养基传代培养制得四种菌悬液,选择卷曲霉素标准品配制测定溶液,分别采用上述菌悬液和五种不同品牌检定培养基进行抗生素微生物检定管碟法试验,考察不同来源菌种、不同品牌传代培养基及不同品牌检定培养基对实验的影响。结果采用相同品牌传代培养基和相同品牌检定培养基进行试验,I号标准菌株生长相对较好;采用相同来源标准菌株和相同品牌检定培养基进行试验,A企业的传代培养基制备的菌悬液菌层生长较好;采用相同来源标准菌株、相同品牌传代培养基和不同品牌检定培养基进行试验,并对检定培养基进行综合评分,结果B企业的检定培养基质量较差,C企业的检定培养基质量较好。结论标准菌株、菌种传代培养基及抗生素检定培养基的来源对效价测定均有影响,而菌种在法定标准中有明确规定,因此在抗生素微生物检定法中应选择最适传代培养基与检定培养基作为效价测定的系统适用性条件;目前国内培养基质量参差不齐,应对培养基质量进行严格控制。

Objective To investigate the in?uence of organism, subculture medium and antibiotic test medium on antibiotic microbiological assay, and determine system suitability criteria for antibiotic microbiological assay. Methods The standard organism ATCC6633 from American type culture collection and the standard organism CMCC (B) 63501 from National Institutes for Food and Drug Control of China were selected to prepare four kinds of suspensions by agar culture medium from A and B enterprises respectively.Capreomycin RS was selected to prepare the test solution, the antibiotic microbial test was carried out with the above suspensions and five different brands of media respectively, in order to investigate the influence of organisms from different sources and different brands of subculture media and verification media. Results Using the same brand subculture medium and the same brand verifcation medium, the growth of standard organism I was relatively good;Using the same standard organisms and the same brand verifcation medium to carry on the experiment, the bacterial suspension layer prepared by enterprise A's subculture medium grew well.Standard organisms from the same source, the same brand culture medium and different brand culture medium were used to carry out the test, and the verifcation medium was comprehensively scored. The results showed that the quality of the verifcation medium of enterprise B was poor, and that of enterprise C was better. Conclusion The source of standard organism, subculture medium and antibiotic test medium all have in?uence on antibiotic microbiological assay, and the organism is clearly stipulated in the statutory standard. Therefore, the best subculture medium and test medium should be selected as the system applicability condition for in microbiological test of antibiotics.At present, the quality of the medium is uneven, and the quality of the medium should be strictly controlled.