OmniLog TM微生物鉴定系统在青海高原野生型鼠疫噬菌体宿主谱检测中的应用研究  

Application of the OmniLog TM microbial identification system in the detection of the host spectrum for wild-type plague phage in Qinghai Plateau

作　　者：李存香[1] 祁芝珍[1] 张青雯[1] 赵海红[1] 马龙 游陪松 杨建国[1] 吴海生[1] 冯建萍[1] LI Cun-xiang;QI Zhi-zhen;ZHANG Qing-wen;ZHAO Hai-hong;MA Long;YOU Pei-song;YANG Jian-guo;WU Hai-sheng;FENG Jian-ping(National Health Commission Co-construction Key Laboratory for Plague Control and Prevention,Qinghai Institute for Endemic Disease Control and Prevention,Xining 811602,China)

机构地区：[1]青海省地方病预防控制所/国家卫生健康委鼠疫防治研究重点实验室(共建),西宁811602

出　　处：《中国人兽共患病学报》2024年第1期21-25,共5页Chinese Journal of Zoonoses

基　　金：青海省医药卫生科技项目(No.2022-wjzd-06)。

摘　　要：目的采用两种微量法检测青海高原野生型鼠疫噬菌体的宿主谱,为今后噬菌体生态学研究和基于宿主范围噬菌体分类研究提供科学依据。方法基于OmniLog TM微生物鉴定系统微量法和微量点滴法,检测3株鼠疫噬菌体对2株鼠疫耶尔森菌(鼠疫疫苗株EV 76、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)的裂解情况。结果476号、087号和072204号鼠疫噬菌体均能成功感染鼠疫疫苗株EV 76、614F并依赖其生长繁殖,且基于OmniLog TM系统33℃培养48 h也能够观察到试验孔中四唑类染料颜色随着噬菌体数量的增加和宿主菌数量的减少而变淡。利用微量点滴法检测3株鼠疫噬菌体对4株非鼠疫耶尔森菌敏感性结果显示,28℃和37℃时476号鼠疫噬菌体对假结核耶尔森菌PTB5敏感,而087号和072204号对其不敏感;476号、087号和072204号鼠疫噬菌体对假结核耶尔森菌PTB3、大肠杆菌V517、小肠结肠炎耶尔森菌52302-2这3株非鼠疫耶尔森菌均不敏感,且基于OmniLog TM系统观察到试验孔与对照孔中细菌生长曲线和四唑类染料颜色分别呈现相似对应的结果。然而,以鼠疫疫苗株EV 76、614F为宿主菌,37℃时072204号鼠疫噬菌体微量点滴于固体培养基上不显示噬菌斑。结论基于OmniLog TM检测系统的噬菌体宿主范围测定法可以作为一种替代传统检测宿主谱的测定方法,在噬菌体与宿主菌相互作用研究中具有较好的应用价值。The growth of three plague phages from Qinghai Plateau in two Yersinia pestis strains(plague vaccine strains EV 76 and 614F)and four non-Yersinia pestis strains(Yersinia pseudotuberculosis PTB3,PTB5,Escherichia coli V517,and Yersinia enterocolitica 52302-2)were detected through a micromethod based on the OmniLog TM microbial identification system and by the drop method,to provide a scientific basis for future ecological studies and classification based on the host range.For plague vaccine strains EV 76 and 614F,successful phage infection and subsequent phage growth were observed in the host bacterium.Diminished bacterial growth and respiration and a concomitant decrease in color were observed with the OmniLog TM microbial identification system at 33℃for 48 h.Yersinia pseudotuberculosis PTB5 was sensitive to Yersinia pestis phage 476,but Yersinia pseudotuberculosis PST5 was insensitive to phage 087 and 072204.Three strains of non-Yersinia pestis(Yersinia pseudotuberculosis PTB3,Escherichia coli V517,and Yersinia enterocolitica 52302-2)were insensitive to Yersinia pestis phages 087,072204,and 476 showed similar growth curves.The growth of phages 476 and 087,as determined with the drop method,in two Yersinia pestis strains(plague vaccine strains EV 76 and 614F)and four non-Yersinia pestis strains(Yersinia pseudotuberculosis PTB3,Escherichia coli V517,and Yersinia enterocolitica 52302-2)showed the same results at 37℃,on the basis of comparisons with the OmniLog TM microbial identification system;in contrast,phages 072204 did not show plaques on solid medium at 37℃with plague vaccine strains EV 76 and 614F.Determination based on the OmniLog TM detection system can be used as an alternative to the traditional determination of the host range,thus providing favorable application value for determining the interaction between the phage and host bacteria.

关 键 词：噬菌体 鼠疫菌 宿主谱 OmniLog TM微生物鉴定系统 点滴法 

分 类 号：R378.61[医药卫生—病原生物学]