微孔板稀释法在活菌培养计数中的应用观察

发布时间：2018-04-13 09:11

本文选题：活菌培养计数 + 孔板微量稀释法　；参考：《中国消毒学杂志》2016年08期

【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。
[Abstract]:Objective to establish a new method for counting live bacteria by using 96 hole plate microdilution instead of traditional test tube dilution. The accuracy and rapidity of the new method were verified by comparing two methods of counting living bacteria.Methods the different blowing times of the new method were compared and the best suction times were selected.In order to verify the accuracy of the new method, the spore suspensions of Bacillus subtilis var. Niger were counted by two living bacteria culture counting methods at the same time.At the same time, the same tube Bacillus subtilis var. Niger spore suspension was diluted by 96 hole plates and counted by drop counting method to compare the error rate of different operators.The neutralizing agent identification test of the same disinfectant was carried out by two methods of living bacteria culture counting to verify the feasibility of applying the new method in the detection of disinfection effect.Results the error rate of different suction times was compared. The best suction times were 20 times. The results of counting spores suspension of Bacillus subtilis var. Niger by 96 hole plate microcount method were consistent with those of traditional method.Using the new method, the error rate between three persons counting a tube of bacteria suspension was 1.69, which was in accordance with the requirements of the 2002 edition of the Technical Specification for Disinfection.And the error rate was much lower than that of the traditional method of counting the live bacteria from Bacillus subtilis var. Niger by traditional methods. In the neutralizer identification test of a disinfectant,The result of the new method is consistent with that of the traditional method.Conclusion the result of the new counting method is consistent with that of the traditional living bacteria count method, and the error rate among different technicians is very small. The new method can be applied to the quantitative germicidal test of disinfectant suspensions.The new culture counting method is more convenient, quick and time saving, which is suitable for further popularization.
【作者单位】：中国人民解放军疾病预防控制所;
【基金】：军队后勤科技计划项目（CWS13C046）
【分类号】：R446.5

【参考文献】