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目的通过四唑鎓盐(XTT)法检测布鲁杆菌疫苗的活菌含量,探讨XTT法快速检测布鲁杆菌活菌含量的可行性。方法将XTT法运用于布鲁杆菌疫苗活菌含量的检测,通过已知活菌浓度的布鲁杆菌液,制备XTT法的吸光值与布鲁杆菌苗活菌含量的关系参比曲线,并根据制备的参比曲线同样运用XTT法检测未知浓度的104M菌液的活菌量,比较传统活菌集落计数(CFU)法与XTT法两者检测布鲁杆菌活菌含量的相关性。结果 XTT法检测布鲁杆菌疫苗的最佳时间应在4 h左右,在一定范围内XTT法所测得的吸光值与细菌活菌含量有较好的相关性,相关系数r=0.997。运用10瓶布鲁杆菌疫苗,通过XTT法与CFU法检测其活菌含量,两者结果相关性较好,Spearman相关系数为0.879,P=0.001。结论 XTT法能在一定活菌浓度范围反应布鲁杆菌疫苗活菌含量,可为快速检测布鲁杆菌活菌含量提供一种新的参考方法。
Objective To detect the viable bacteria content of Brucella vaccine by tetrazolium salt (XTT) method and to explore the feasibility of rapid detection of viable bacteria content of Brucella by XTT method. Methods The XTT method was applied to the detection of viable bacteria in Brucella vaccine, the Brucella bacterium with known concentration of live bacteria was used to prepare the reference curve of the relationship between the absorbance of XTT method and the content of live bacteria of Brucella, The reference curve prepared also used the XTT method to detect the viable cell count of the 104M bacterial solution with unknown concentration. The correlation between the viable count of viable bacteria of Brucella was compared between the traditional viable colony count (CFU) method and the XTT method. Results The optimum time for detecting Brucella vaccine by XTT method should be about 4 h. There was a good correlation between the absorbance measured by XTT method and the content of viable bacteria in a certain range. The correlation coefficient was 0.997. The results of Spearman’s correlation coefficient were 0.879, P = 0.001 by using 10 Brucella vaccine and detecting the viable cell viability by XTT method and CFU method. Conclusion The XTT method can reflect the content of viable Brucella vaccine in a range of viable bacteria, which can provide a new reference method for rapid detection of viable bacteria in Brucella.