基于微量液体培养基间接评估鼠疫噬菌体生长方法的建立  

Establishment of methods for indirectly assessing Yersinia pestis Bacteriophage growth based on microscale liquid medium

作　　者：李存香[1] 赵海红[1] 张青雯[1] 金星[1] 辛文媛 张丽 冯建萍[1] 金泳[1] 代瑞霞[1] 祁芝珍[1] LI Cunxiang;ZHAO Haihong;ZHANG Qingwen;JIN Xing;XIN Wenyuan;ZHANG Li;FENG Jianping;JIN Yong;DAI Ruixia;QI Zhizhen(Qinghai Institute for Endemic Disease Control and Prevention,Xining 810021,China)

机构地区：[1]青海省地方病预防控制所,西宁810021

出　　处：《医学动物防制》2023年第5期419-422,共4页Journal of Medical Pest Control

基　　金：中央级公益性科研院所基本科研项目(2019PT310003);青海省鼠疫防控及研究重点实验室(2021-ZJ-Y15);青海省医药卫生科技项目指导性计划课题(2020-wjzdx-110)。

摘　　要：目的利用微量液体培养基建立评估鼠疫噬菌体生长的方法,为今后鼠疫噬菌体的分离、宿主谱鉴定、生物学特性研究等提供技术依据。方法利用OmniLog TM微生物鉴定系统,检测微生物细胞对不同碳源呼吸代谢导致的显色反应和微生物生长造成浊度差异的原理,采用微量液体培养法检测诊断用鼠疫噬菌体在2株鼠疫耶尔森菌(鼠疫弱毒菌EV76、614F)和4株非鼠疫耶尔森菌(假结核耶尔森菌PTB3、PTB5、大肠杆菌V517、小肠结肠炎耶尔森菌52302-2)的生长情况。结果实验菌株及诊断用鼠疫噬菌体混合物经OmniLog TM微生物鉴定系统33℃作用48h,以鼠疫疫苗株EV76、鼠疫弱毒菌614F为试验菌,试验行孔的生长曲线呈一条横线,且细菌浓度对数值不超过100;四唑类染料的颜色随着噬菌体数量的减少和宿主菌数量的增多逐渐加深,表明诊断用鼠疫噬菌体能感染鼠疫弱毒菌EV76和614F。以假结核耶尔森菌PTB3、PTB5、大肠杆菌V517、小肠结肠炎耶尔森菌52302-24株非鼠疫耶尔森菌为试验菌,每行6孔均呈现几乎相同的对应试验菌株的S型生长曲线,说明诊断用鼠疫噬菌体不能感染对应行的试验菌株。利用OmniLog TM微生物鉴定系统检测诊断用鼠疫噬菌体对6株试验菌株的灵敏度,结果与液体培养基和半固体培养基的常规检测结果一致。结论成功建立基于OmniLog TM微生物鉴定系统的微量液体培养基间接检测鼠疫噬菌体生长的方法,评估鼠疫噬菌体对宿主菌的敏感度,是一种经济、灵敏、准确的鉴定方法。Objective A method to evaluate the growth of Yersinia pestis Bacteriophage was established by using a microscale liquid medium,which provided a technical basis for the isolation of phage,host spectrum identification,and biological characteristics in the future.Methods The sensitivity of Yersinia pestis Bacteriophage to plague vaccine strain EV76 was detected according to the principle of respiration metabolism of different carbon sources and specific chromogenic results of microbial cells by using OmniLog TM microbial identification system.The strains include Yersinia pestis EV76,Yersinia pestis 614F,Yersinia pseudotuberculosis PTB3,Yersinia pseudotuberculosis PTB5,Escherichia coli V517,and Yersinia enterocolitica 52302-2.Results The mixture of the test strain and the diagnostic Yersinia pestis Bacteriophage was reacted by the OmniLog TM microbial identification system at 33℃for 48 hours,and the plague vaccine strain EV76 and plague attenuated bacterium 614F were used as the test bacteria,and the growth curve of the test row wells was a horizontal line,and the logarithmic bacterial concentration did not exceed 100.The color of tetrazolium dyes gradually deepened with the decrease of the number of bacteriophages and the increase of the number of host bacteria,indicating that the diagnostic Yersinia pestis Bacteriophage could infect the plague attenuated bacteria EV76 and 614F.Four non-Yersinia pestis(Yersinia pseudotuberculosis PTB3 and PTB5,Escherichia coli V517,and Yersinia enterocolitica 52302-2)were test bacteria,and the six holes in each row showed almost identical S-shaped growth curves of corresponding test strains,indicating that the diagnostic Yersinia pestis Bacteriophage could not infect the corresponding test strains.The sensitivity of the diagnostic Yersinia pestis Bacteriophage to the six test strains was detected by the OmniLog TM microbial identification system,and the results were consistent with the conventional detection results of liquid medium and semi-solid medium.Conclusion The method of i

关 键 词：鼠疫菌 噬菌体 微量液体培养基 OmniLog TM微生物鉴定系统 

分 类 号：R378.60[医药卫生—病原生物学]