贝氏柯克斯体耐受渗透脱水研究模型的建立  

Establishment of a research model for the osmotic dehydration tolerance of Coxiella burnetii

作　　者：王涛 孙静 尹白露 于永慧 何卫平 段学章 WANG Tao;SUN Jing;YIN Bai-lu;YU Yong-hui;HE Wei-ping;DUAN Xue-zhang(Department of Radiation Oncology,Division of medical Oncology,The Fifth Medical Center of PLA General Hospital,Beijing 100039,China;Senior Department of Infectious Disease,The Fifth Medical Center of PLA General Hospital,Beijing 100039,China;State Key Laboratory of Pathogen and Biosecurity,Beijing Institute of Microbiology and Epidemiology,Beijing 100071,China)

机构地区：[1]解放军总医院第五医学中心肿瘤医学部放射治疗科,北京100039 [2]解放军总医院第五医学中心感染病医学部,国家感染临床医学研究中心,北京100039 [3]军事医学研究院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《中国人兽共患病学报》2024年第5期415-420,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金青年项目(No.82003211)。

摘　　要：目的基于无生命培养体系,探索建立贝氏柯克斯体耐受脱水的实验室研究模型。方法梯度调整贝氏柯克斯体无生命培养体系中的渗透压条件,利用qPCR定量测定不同渗透压下贝氏柯克斯体在培养周期内基因组拷贝数变化并描记生长曲线,进一步利用相差显微镜、透射电子显微镜对真核细胞以及无细胞培养方法所得贝氏柯克斯体进行形态学分析,并感染BGMK细胞测定不同渗透压培养所得贝氏柯克斯体的感染性VFU。结果贝氏柯克斯体可在高渗透压条件的无生命培养基中适应性生存7 d以上,经高渗透压培养后的菌体脱水收缩且大小类似于小贝氏体,同时感染后24 h的VFU较对照组降低。结论成功建立了贝氏柯克斯体耐受高渗压脱水的研究模型,为下一步用蛋白质组学等方法研究贝氏柯克斯体在自然环境中耐受干燥脱水的遗传学机制奠定了基础。The aim of this study was to establish a laboratory research model for the desiccation tolerance of Coxiella burnetii(C.burnetii),based on an axenic culture system.The conditions for osmotic pressure in the axenic culture system of C.burnetii were set via a gradient.Quantitative PCR was used to determine the C.burnetii genome equivalents during the culture cycle under different osmotic pressures,and the growth curves were recorded.In addition,the bacterial manifestations of C.burnetii obtained from eukaryotic cell cultures or cell-free cultures were analyzed with phase contrast microscopy and transmission electron microscopy(TEM).The bacterial infection levels and vacuole forming units(VFU) were measured by infection of BGMK cells.C.burnetii showed as many as 7 days of adaptive survival in osmotic axenic medium under high osmotic conditions.The bacteria shrank by dehydration under extremely high osmotic pressure and appeared primarily as hypo-hydrated small cell variants(SCVs).The VFUs were significantly diminished 24 hours after infection,as compared with the parallel contrasts.The method for researching desiccation tolerance was thus successfully established.This method provides a basis for further investigation of the genetic mechanisms of the anti-desiccation properties of C.burnetii in the natural environment,through proteomics and other methods.

关 键 词：贝氏柯克斯体 Q热 渗透压 脱水 无生命培养 

分 类 号：R383.3[医药卫生—医学寄生虫学]