微孔板稀释法在活菌培养计数中的应用观察  

Application observation of viable organism cultivation through the dilution method of microwell plate

作　　者：韩杰[1] 饶林[1] 饶亚岚[1] 苏裕心[1] 魏秋华[1] 姚楚水[1] 

机构地区：[1]中国人民解放军疾病预防控制所,北京100071

出　　处：《中国消毒学杂志》2016年第8期728-731,共4页Chinese Journal of Disinfection

基　　金：军队后勤科技计划项目(CWS13C046)

摘　　要：目的以96孔板微量稀释法代替传统试管稀释法建立了新的活菌计数方法(也简称新方法或96孔板计数法),通过两种活菌培养计数方法的比较,验证新方法的准确性、快捷性。方法对新方法的不同吹吸次数进行比较,选择最佳的吹吸次数。用两种活菌培养计数方法同时对同一管枯草杆菌黑色变种芽孢悬液进行计数,验证新方法的准确性;不同实验技术人员同时对同一管枯草杆菌黑色变种芽孢悬液用96孔板进行系列稀释,用滴液计数法计数,比较不同人员操作的误差率;用两种活菌培养计数方法对同一种消毒剂进行中和剂鉴定试验,验证新方法在消毒效果检测中应用的可行性。结果经不同吹吸次数的误差率比较,20次为最佳吹吸次数;96孔板微量计数法对枯草杆菌黑色变种芽孢悬液计数结果与传统方法计数结果一致;3名不同实验技术人员采用新方法同时对一管菌悬液计数,3人之间的误差率为1.69%,符合2002年版《消毒技术规范》要求,且误差率远低于新入职实验技术人员用传统方法对枯草杆菌黑色变种芽孢悬液活菌培养计数结果;在一种消毒剂的中和剂鉴定试验中,新方法与传统方法活菌培养计数结果一致。结论新的活菌计数法计数结果与传统活菌培养计数结果一致,且不同实验技术人员之间的误差率很小,新的培养计数方法可以应用到消毒剂悬液定量杀菌试验中,且采用新的培养计数方法更方便、快捷,节省时间,对实验人员技术要求不高,适合进一步推广。Objective To establish the new method of viable count by the 96 pore plate microdilution method instead the tradition tube dilution method（also to abbreviate new method or 96 pore plate counting method ） ,through compare the two methods of viable count, to validate the new methods accuracy and shortcut. Methods Through compare the distinct blow and absorb frequency, to choose the best number of times. Using the same suspensions of spore to count by two methods of viable count, to validate the accuracy of new method;with the purpose of comparing error ration of operate by different per- sonnel, let different experiment technologist use the same suspensions of spore to viable count, with 96 pore plate in progress series dilution, to count by drip counting method. To validate the feasibility of new method in the detection of disinfection re- sult,with neutralizer qualification test for one disinfectant by two methods. Results Through the comparison of different blow and absorb frequency, the twentieth times best ; the counting result of new method and tradition method is coincidence ; three technologists use the new method to count,that the error rate is 1.69% ,which consistent with the〈〈Disinfection Tech- nology Regulation）〉of 2002 edition, and lower than the latest technologists who use the tradition method;The two methods＇re- sult is the same in the neutralizer qualification test of one antiseptic. Conclusion Through the comparison of the twe meth- ods, the result with one accord, and the error ration is low in different experiment technologist, the new method could be used in the suspension quantitative germicidal, it is more convenient and quickly, saving time, the specification is not high for ex- perimenter, so it is worth to further generalization.

关 键 词：活菌培养计数 96孔板微量稀释法 传统稀释法 比较 

分 类 号：Q331[生物学—遗传学] R187[医药卫生—流行病学]