基于响应面分析的高抗原活性猪水肿病大肠杆菌疫苗培养基的研制及效果评价  

Development and effect evaluation of a vaccine culture medium with high antigenic activity for the prevention of porcine edema disease caused by Escherichia coli based on response surface methodology

作　　者：张阳阳 万强 牛得权 苗森 蒋富凤 刘翔 曲晓莹 张菊梅 蔡芷荷 陈博 吴清平 ZHANG Yangyang;WAN Qiang;NIU Dequan;MIAO Sen;JIANG Fufeng;LIU Xiang;QU Xiaoying;ZHANG Jumei;CAI Zhihe;CHEN Bo;WU Qingping(College of Food and Biological Engineering,Shaanxi University of Science and Technology,Xi’an 710021,Shaanxi,China;State Key Laboratory of Applied Microbiology Southern China,Guangdong Provincial Key Laboratory of Microbial Safety and Health,Key Laboratory of Agricultural Microbiomics and Precision Application,Ministry of Agriculture and Rural Affairs,Institute of Microbiology,Guangdong Academy of Sciences,Guangzhou 510070,Guangdong,China;Guangdong Huankai Microbial Limited Company,Zhaoqing 526238,Guangdong,China)

机构地区：[1]陕西科技大学食品与生物工程学院,陕西西安710021 [2]广东省科学院微生物研究所、华南应用微生物国家重点实验室、广东省微生物安全与健康重点实验室农业农村部农业微生物组学与精准应用重点实验室,广东广州510070 [3]广东环凯生物科技有限公司,广东肇庆526238

出　　处：《微生物学通报》2023年第9期4141-4153,共13页Microbiology China

基　　金：国家自然科学基金(32072325);广东省重点研发计划(2022B1111040002);广东省科学院创新发展专项(2020GDASYL-20200103024)。

摘　　要：【背景】猪水肿病大肠杆菌引发的疾病造成了很大的危害,但现有培养基存在培养密度低的问题。【目的】研制出高抗原活性猪水肿病大肠杆菌疫苗培养基。【方法】以常用的市售猪水肿培养基为对照,通过单因素试验、爬坡试验(Plackett-Burman,PB)、响应面(Box-Behnken,BB)试验对猪水肿培养基进行响应面优化,得到猪水肿培养基最优配方。以响应面试验得到的培养基培养猪水肿病大肠杆菌,评价不同培养时间点菌株的抗原活性,制作灭活疫苗,进行动物免疫保护试验。【结果】对研制的培养基进行扩大培养验证,发现扩大培养得到的菌株活菌数可达5×10^(9) CFU/mL以上,约为对照组的2倍。制备的灭活疫苗效价可达1:140000,并在9 h时抗原蛋白效价达到最高。【结论】本研究研制出的疫苗培养基显著提高了猪大肠杆菌菌体密度,并可提高菌体抗原活性,为猪水肿病灭活疫苗的制备提供了技术指引。[Background]Porcine edema disease caused by Escherichia coli is a major disease attacking pigs.The existing culture medium used for vaccine preparation has a low cell density.[Objective]To develop a vaccine culture medium with high antigenic activity for the prevention of porcine edema disease.[Methods]A commercially available vaccine culture medium against porcine edema disease was used as the control.The response surface methodology was employed to optimize the composition of the culture medium through single-factor experiments,Plackett-Burman(PB)design,and Box-Behnken(BB)design.The optimized culture medium was used to culture the E.coli eliciting porcine edema disease,and the antigenic activity of the culture at different time points was evaluated.In addition,the inactivated vaccine was prepared and used for animal immunization.[Results]The expanded culture of the pathogen in the developed culture medium showed that the viable cell count of the strain reached more than 5×10^(9) CFU/mL,approximately twice that of the control group.Meanwhile,the titer of the inactivated vaccine reached 1:140000,and the antigen protein titer peaked at the time point of 9 h.[Conclusion]The developed vaccine culture medium significantly improved the cell density and antigenic activity of E.coli causing porcine edema disease,providing technical guidance for preparing inactivated vaccines against porcine edema disease.

关 键 词：猪水肿病 响应面法 疫苗培养基 高抗原活性 

分 类 号：S858.28[农业科学—临床兽医学]