对《硫乙醇酸盐流体培养基灵敏度检查(草案)》的验证试验  

Verification of the Sensitivity Examination Protocol on Thioglycollate Fluid Culture Medium

作　　者：高尚先[1] 孙彬裕[1] 康国华[1] 刘艳[1] 曲守方[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《药物分析杂志》2005年第7期849-851,共3页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:对新提出的《硫乙醇酸盐流体培养基灵敏度检查(草案)》的准确性、可操作性和可重复性进行验证。方法:将质控菌乙型溶血性链球菌、生孢子梭状芽孢杆菌(简称生孢梭菌)、短芽孢杆菌接种于适宜的培养基培养,取新鲜培养物用0.9% 无菌氯化钠制成浊度稍高于标准比浊管的均匀菌液,然后分别用0.9%无菌氯化钠溶液进行系列稀释至约相当于10-100 CFU·mL-1,作为工作菌液。取上述工作菌液分别接种于3管(1 mL·管-1)硫乙醇酸盐流体培养基中,置35℃培养72h(短芽孢杆菌培养5 d)。同时将上述工作菌液进行CFU·mL-1计数:将乙型溶血性链球菌和短芽孢杆菌采用涂抹法分别接种于血琼脂培养基和营养琼脂培养基,置有氧条件35℃培养24h,生孢梭菌采用浇注法接种于生孢梭菌CFU计数培养基,置厌氧条件35℃培养18-20 h,用未接种的培养基作为空白对照。结果:当上述3种质控菌工作菌液的平皿CFU计数在10-100 CFU·mL-1时,其接种的3管硫乙醇酸盐流体培养基均呈现生长。结论:新提出的《硫乙醇酸盐流体培养基灵敏度检查(草案)》准确性、可操作性和重复性良好,易于推广和实施。可以提出作为对中国药典(2005年版)中硫乙醇酸盐流体培养基灵敏度检查的修订草案。Objective:To verify the accuracy, maneuverability and repeatability of the sensitivity examination protocol on the thioglycollate fluid culture medium, which is put forward newly. Method:As the quality control aerobes, Streptococcus B - hemolyticus and Bacillus brevis were inoculated on the blood agar slant and the nutrient agar slant,while the quality control anaerobes,Clostridiums sporogenes was inoculated on the thioglycollate liquid culture medium(without agar) for enrichment. The fresh culture was put into the test tube and diluted with 0.9% sterilized saltwater to the concentration of 10-100 CFU·mL-1,from which we got different working bacterial suspension. Each of the different working suspension was inoculated into three test tubes containing 9 mL thioglycollate fluid culture medium(1 mL per tube). The tubes with Streptococcus B-hemolyticus and Bacillus brevis were put under aerobic condition at 35℃ for 72 h and five days respectively, while the tubes with Clostridium sporogenes were put under anaerobic condition at 35℃ for 72 h. At the same time, we carry out the CFU counting test by inoculating 1 mL working suspendsion of Streptococcus B - hemolyticus onto the blood agar plate, 1 mL working suspendsion of Bacillus brevis onto the nutrient agar plate using L stick daubing method, and 1 mL working suspendsion of Clostridium sporogenes onto the CFU counting culture medium plate using poling method. The former two kind of plates were put under aerobic condition at 35℃for 24 h, while latter kind of plate was put under anaerobic condition at 35℃ for 18-20 h. Uninoculated medium is used as control. Results:When the CFU counting we got from the plates ranging from 10 to 100 CFU·mL-1, the three kind of quality control strains inoculated in the three paralled liquid thioglycollate culture medium grow well. Conclusion:This study led to the conclusion that the accuracy, maneuverability and repeatability of the sensitivity examination protocol on the thioglycollate fluid culture medium is quite well and ea

关 键 词：硫乙醇酸盐 灵敏度 流体 检查 验证试验 乙型溶血性链球菌 短芽孢杆菌 营养琼脂培养基 生孢梭菌 梭状芽孢杆菌 血琼脂培养基 2005年版 可操作性 氯化钠溶液 可重复性 空白对照 中国药典 准确性 质控菌 CFU 菌液 培养物 

分 类 号：R927.12[医药卫生—药学] R651.150.2