基于流式细胞仪测定细菌杀菌率方法研究  

Study on determination of bactericidal rate based on flowcytometry

作　　者：刘中华[1] 卢洁元 赵巍薇 翟平平 李睿[1] LIU Zhong-hua;LU Jie-yuan;ZHAO Wei-wei;ZHAI Ping-ping;LI Rui(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China;Jiangxi Institute of Food Inspection and Testing,Nanchang 330000,China)

机构地区：[1]武汉轻工大学生命科学与技术学院,武汉430023 [2]江西省食品检验检测研究院,南昌330000

出　　处：《武汉轻工大学学报》2021年第5期40-44,共5页Journal of Wuhan Polytechnic University

基　　金：江西省市场监督管理局科技计划项目(GSJK202104).

摘　　要：传统的细菌杀菌率测定方法耗时费力,且不能够精确地检测到处于休眠状态的菌体。探讨了使用流式细胞仪测定杀菌率的可行性和准确性。使用10.24 mg/mL氨苄西林抗生素溶液处理奇异变形杆菌012,分别采用流式细胞仪和平板菌落计数法测定细菌数目并绘制杀菌曲线。结果显示,流式细胞仪测定死细胞数占圈门细胞数随培养时间的延长而升高,24 h后杀菌率为18.28%;PCA平板菌落计数所得杀菌率均>99%,杀菌率曲线自6 h之后基本保持稳定。检测结果显示两种方法所测得的杀菌率曲线不能很好地拟合,主要是因为流式细胞仪可检测细胞膜受损的细胞及处于休眠状态的菌体,而这些细菌无法通过传统培养方法计数。结果表明流式细胞仪和平板菌落计数结合,可测定休眠态细菌,全面了解杀菌效率。Traditional plate culture counting is time-consuming and laborious and unable to detect the dormant bacteria.The potential usage of flow cytometry for bactericidal rate was measured in this paper.Proteus mirabilis 012 was treated with 10.24 mg/mL of ampicillin.Then flow cytometry and plate culture counting were used to determine the antibiotic killing curve.The results showed that the number of dead cells increased with the extension of culture time,and the percentage of dead cells was recorded to be 18.28%of the initial cell count after 12 h exposure.The bactericidal rate obtained from PCA plate colony counting was above 99%and maintained stability after 6 h.The antibiotic killing curves measured by the two approaches could not be consistent.It may due to the dormancy cells and membrane-damaged cells.These cells were not detected by plate counting.However,they could be detected by flow cytometry.The combination of flow cytometry and plate counting will be accurate as dormancy cells are included in the overall of measured cells.

关 键 词：流式细胞仪 氨苄西林 杀菌率 平板菌落计数 

分 类 号：Q93[生物学—微生物学]