不同方法对多房棘球绦虫虫卵破碎效果的镜下观察研究

The Effect of Different Methods for the Crushing of Echinococcus Multilocularis Eggs

目的通过镜下观察、计数,比较不同方法对多房棘球绦虫虫卵的破碎效果。方法以液氮-95℃水浴反复冻融、裂解液消化、钢珠震荡和超声破碎4种方法在不同条件下对多房棘球绦虫虫卵进行处理,镜下观察虫卵破碎情况,并根据虫卵破碎程度进行分级,通过计数不同分级条件下虫卵数量,比较、分析不同方法对棘球绦虫卵的破碎效果。结果 4种方法对多房棘球绦虫虫卵均有一定的破碎效果,钢珠震荡和超声破碎对虫卵的作用效果优于反复冻融和裂解液消化。钢珠震荡破碎作用3 min和200 W条件下的超声作用可使检测虫卵基本全部破碎。结论钢珠震荡和超声裂解方法对多房棘球绦虫虫卵破碎处理效果较好,具有作为棘球绦虫粪-PCR检测预处理的应用潜力。

Objective To compare crushing effects of different methods for Echinococcus multilocularis eggs based on microscopic observation and counting of the number of broken eggs.Methods Echinococcus multilocularis eggs were treated by four methods under different conditions,including repeated liquid nitrogen-95℃ water bath,lysate digestion,disruption using bead mills and ultrasound.The crushing status of the eggs was observed by using the microscope.The number of crushed eggs was counted,and the crushing effect of those four methods was compared.Results Those four methods have certain effects on breaking of the eggs.The effect of disruption using bead mills and ultrasound were better than that of the other two methods.Disruption using bead mills for 3 minutes and ultrasound under the condition of 200 W can almost break all detected eggs.Conclusion Disruption using bead mills and ultrasound is effective to crush the Echinococcus eggs.These two methods have a potential to be a pretreatment method for the molecular detection of Echinococcus eggs in fecal samples.