人芽囊原虫保存方法的初步研究  

A Preliminary Study on Preservation Methods of Blastocystis hominis

作　　者：田春林[1] 万孝玲[1] 刘登宇[1] 何登贤[1] 

机构地区：[1]广西医科大学寄生虫学教研室,南宁530021

出　　处：《中国寄生虫学与寄生虫病杂志》2007年第1期57-60,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：广西青年科学基金(No.0542046)~~

摘　　要：目的观察冻存剂、温度等因素对人芽囊原虫活力的影响,探索理想的人芽囊原虫保存方法。方法从患者阳性粪便中分离人芽囊原虫,分装到2 ml无菌冻存管内,分别加入10%二甲基亚砜(dimethylsulfoxide,DMSO)、40%丙三醇(GL)和15%乙二醇(EG)作冻存剂,放置于不同的温度下保存,用台盼蓝染色法和原虫培养法测定细胞的活力和增殖能力。结果虫体在室温(18℃～20℃)下可存活3周,在4℃～6℃存活不到1周。加入冻存剂后,置-20℃和液氮(-196℃)下可存活3个月以上。用40%GL作冻存剂,于液氮低温下冻存的虫体保存半年后复苏,活力仍达41.7%,培养72 h后多见分裂相细胞。结论应用40%GL作为冻存剂,在液氮中低温保存人芽囊原虫效果较佳。4℃不适于保存人芽囊原虫。Objective To observe the effect of different cryoprotective agents and temperature factors on the viability of Blastocystis hominis so as to explore the ideal method for preservation of B. hominis. Methods B. hominis agents were obtained from a patient＇s fecal specimen. Having washed by normal saline and divided into tubes, the samples were cryopreserved in -20 ℃ refrigerator or in -196 ℃ liquid nitrogen with 10% DMSO, 40% glycerol and 15% ethylene glycol respectively. The thawed B. hominis agents were then used for culture. By trypan blue staining and microscopy, the viability and proliferation of those resuscitative cells were investigated. Results B. hominis survived for 3 weeks at 18 %-20 ℃ while less than 1 week at 4 %-6 %. When stored in -20 ℃ refrigerator or liquid nitrogen with cryoprotective agents, they survived for more than 3 months. The cryopreservation with 40% glycerol at -196 ℃ for 6 months resulted in 41.7% viability of the revivified cells. Cleavage cells were easily observed after culturing for 72 hours. Conclusion Preserving B. hominis in liquid nitrogen with 40% glycerol is an optimal cryopreservation protocol.

关 键 词：人芽囊原虫 保存 温度 

分 类 号：R382.9[医药卫生—医学寄生虫学]