布鲁菌Tb噬菌体效价噬斑测定法的建立  

Establishment of a plaque assay method for determination of phage Tb titer for Brucella

作　　者：张园园 付丽丽[1] 赵爱华[1] 魏东[1] Zhang Yuanyuan;Fu Lili;Zhao Aihua;Wei Dong(Division of Tuberculosis Vaccines and Allergen,National Institute for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院结核病疫苗和过敏原产品室,北京102629

出　　处：《国际生物制品学杂志》2022年第3期159-162,共4页International Journal of Biologicals

摘　　要：目的建立标准化的布鲁菌Tb噬菌体效价噬斑测定方法。方法通过比较单层琼脂平板法与双层琼脂平板法,分析宿主菌接种浓度、吸附时间及培养时间等参数,标准化布鲁菌Tb噬菌体效价测定方法,并分析其精密性。结果虽然两种方法测定的噬菌体效价结果无差别,但双层琼脂平板法的噬斑背景更清晰,易于结果观察。经优化后确定布鲁菌Tb噬菌体效价测定条件:宿主菌的最适接种浓度为1.0×10^(9) ml^(-1),宿主菌与噬菌体混合后不需吸附,培养3~5 d后进行噬斑计数。建立的标准检测方法具有较好的精密性。结论建立了布鲁菌Tb噬菌体效价噬斑测定方法,为布鲁菌Tb噬菌体及布鲁菌活疫苗质量控制奠定了基础。Objective To establish a plaque assay method in determination of phage Tb titer for Brucella.Methods Single-layer agar plate method and double-layer agar plate method were compared.A plaque assay method for determination of titer of phage for Brucella was developed by optimizing the concentration of Brucella cells for inoculation,adsorption time and incubation time.Precision for the developed method was estimated.Results Although the phage titer results by both methods were similar,the plaque background of double-layer agar plate method was more clear and easy to observe.The conditions of optimized method for Brucella phage Tb titer measurement were determined.The optimal Brucella cell concentration for inoculation was 1.0×10^(9) ml^(-1).Adsorption after the mixture of host and phage was not necessary.Plaques were counted after 3-5 d culture.It showed a good precision for the developed method.Conclusion A plaque assay method in determination of phage Tb titer for Brucella is developed,laying foundation for quality control of phage Tb for Brucella and brucellosis vaccine.

关 键 词：布鲁菌 噬菌体 效价 常规裂解稀释度 

分 类 号：R516.7[医药卫生—内科学]