分光光度法测定金黄色葡萄球菌菌液浓度方法的建立  

Establishment of Concentration Determination of Staphylococcus aureus by Spectrophotometry

作　　者：肖敏[1] 杨峰[1] 王旭荣[1] 罗金印[1] 李新圃[1] 陈炅然[1] 李宏胜[1] 

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所、农业部兽用药物创制重点实验室、甘肃省新兽药工程重点实验室,甘肃兰州730050

出　　处：《动物医学进展》2014年第11期40-43,共4页Progress In Veterinary Medicine

基　　金：“十二五”国家科技支撑计划(2012BAD12B03);甘肃省农业科技创新项目(GNCX-2013-59);甘肃省农业生物技术研究与应用开发项目(GNSW-2013-28)

摘　　要：为了建立一种通过吸光度测定,能够快速准确检测奶牛乳房炎多联苗中金黄色葡萄球菌抗原浓度的方法。用血平板培养至生长高峰期的金黄色葡萄球菌用生理盐水将菌苔洗下,稀释成不同浓度的细菌悬浮液,然后用平板计数法测定不同浓度菌液的活菌数,同时在波长450nm条件下测定其相应的吸光度,进而研究两者之间的相关性。结果表明,金黄色葡萄球菌的细菌数与其吸光度呈直线线性相关。建立了金黄色葡萄球菌菌液浓度与吸光度之间的回归方程式y=8.410 6x-0.272 6(R2=0.975 8)。通过测定细菌悬浮液的吸光度,可以快速计算出该菌的菌液浓度。为了验证该方法的准确性,通过平板培养了4批金黄色葡萄球菌菌液,进行了传统平板计数方法和吸光度法的比较和验证,证明吸光度测定法更为简单、快速和准确,且优于平板计数法,可以作为奶牛乳房炎多联苗中金黄色葡萄球菌抗原浓度的测定方法。This study aimed to establish a method to measure the Staphylococcus aureus antigen concentra-tion in multiple vaccine involved in bovine mastitis.Staphylococcus aureus colony in blood agar plate was washed down and diluted to different bacterial suspension.The bacterial counts and absorbance （A450） were determined by plate count method and spectrophotometer method,respectively.The correlation be-tween bacterial counts and absorbance （A450）was studied.The result suggested that Staphylococcus au-reus counts and absorbency showed linear correlation.a regression equation was established between Staphylococcus aureus bacterial suspension concentration and absorbancy （y=8.410 6x-0.272 6,R2 =0.975 8）.The bacterial concentration can be calculated quickly by measuring the absorbancy of Staphylo-coccus aureus .Four batch of Staphylococcus aureus were used to compare and verify the plate count meth-od and spectrophotometer method,which showed that the spectrophotometer method was more simple, quick,and exact,and batter than that of plate count method,suggesting that it can be used to detect Staphylococcus aureus antigen concentration in multiple vaccine involved in bovine mastitis.

关 键 词：奶牛乳房炎 金黄色葡萄球菌 活菌数 吸光度 

分 类 号：S852.611[农业科学—基础兽医学]