结核分枝杆菌菌悬液光密度值与菌落计数的关联性

发布时间：2018-04-16 15:18

本文选题：结核分枝杆菌 + 菌落形成单位　；参考：《中国感染控制杂志》2016年03期

【摘要】：目的建立一种基于光密度值计算结核分枝杆菌菌落数的可靠方法。方法利用低频超声和玻璃珠研磨两种方法制备H37Ra菌悬液,菌悬液2倍梯度稀释后,分别测定各个稀释度菌悬液在600 nm处的光密度值(OD600值),并分析OD600值与稀释倍数曲线,确定最佳的菌悬液制备方法,OD600线性范围,以及OD600值与CFU关联曲线。结果 OD600值为0.1~0.6,线性范围内OD600值与稀释倍数线性回归分析结果显示,玻璃珠研磨法和低频超声法相关系数(R2)分别为0.98、1.00,均呈良好的相关性,且低频超声法比玻璃珠研磨法的相关性好,其菌液分散更均匀。OD600值与CFU值线性回归分析结果显示,玻璃珠研磨法和低频超声法回归方程分别是:CFU=2.35×107×OD600+4.42×105、CFU=3.26×107×OD600+6.89×105。结论低频超声法是一种较好的结核分枝杆菌菌悬液制备方法,结合OD600值测定,可成为一种可靠、快速的结核分枝杆菌定量方法。
[Abstract]:Objective to establish a reliable method for calculating the colony number of Mycobacterium tuberculosis based on optical density.Methods the bacterial suspension of H37Ra was prepared by low frequency ultrasound and glass bead grinding. After the bacterial suspension was diluted by 2 times gradient, the optical density of each diluent was measured at 600nm and the value of OD600 was measured, and the curves of OD600 value and dilution multiple were analyzed.The linear range of the optimal method for preparation of bacteria suspension was determined, and the correlation curve between OD600 value and CFU was also determined.Results the OD600 value was 0. 1 ~ 0. 6. The linear regression analysis between OD600 value and dilution multiple showed that the correlation coefficient of glass bead grinding method and low frequency ultrasonic method was 0. 98% and 1. 00, respectively, showing a good correlation.The correlation between low frequency ultrasonic method and glass ball grinding method was better than that of glass ball grinding method. The linear regression analysis showed that the regression equations of glass bead grinding method and low frequency ultrasonic method were 3.26 脳 107 脳 OD600 6.89 脳 10 5 and 2.35 脳 10 7 脳 10 5 脳 OD600 4. 42 脳 10 5 脳 10 5 OD600 respectively.Conclusion the low frequency ultrasonic method is a good method for the preparation of Mycobacterium tuberculosis suspensions. Combined with the determination of OD600 value, it can be a reliable and rapid quantitative method for Mycobacterium tuberculosis.
【作者单位】：复旦大学生命科学学院;
【基金】：上海市科技支撑项目(15431900200)
【分类号】：R378.911

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