高纯度日本血吸虫卵的分离及活性鉴定  

Isolation and identification of viability of Schistosoma japonicum eggs with high purity

作　　者：钟政荣[1] 徐云侠[2] 宫继勇[1] 罗庆礼[3] 闻慧琴[3] 宋晓蓉[3] 沈继龙[3] 

机构地区：[1]安徽省蚌埠医学院第一附属医院检验科,蚌埠233004 [2]安徽省蚌埠医学院第一附属医院神经内科,蚌埠233004 [3]安徽医科大学病原生物学教研室,安徽省病原生物学省级实验室,安徽省人兽共患病重点实验室,合肥230032

出　　处：《中国人兽共患病学报》2012年第3期209-212,共4页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.81071376);安徽省教育厅自然科学基金重点项目(No.KJ2011A205);安徽省自然科学基金(No.090413089)

摘　　要：目的探讨制备高纯度、高生物活性的的日本血吸虫卵分离方法。方法将匀浆后的血吸虫感染的兔肝组织悬液分级过滤,再将滤液离心,去除组织碎片,分离的虫卵再次经过325目分样筛过滤,收集筛上虫卵,最后将收集的虫卵用密度1.070的Percoll分离液梯度离心,吸取下层Percoll液中的纯化虫卵;显微镜观察分离虫卵的纯度,吖啶橙染色观察虫卵的死活,环卵沉淀试验鉴定虫卵的分泌活性。结果每个肝脏获得约1g左右的虫卵,分离的虫卵纯度高,吖啶橙染色和环卵沉淀试验结果显示,分离的活虫卵比例高、生物活性好。结论该虫卵分离方法操作简单、用时短、效果好,分离的虫卵完全满足各种研究的需要。To modify the isolation methods of Schistosoma japonicum eggs with high purity and high biological activity,the liver tissues from rabbits 6 weeks post-infection were homogenizated before the resulting suspensions were consecutively fractional filtration.The filtrate was centrifuged,the supernatant fluid and tissue-containing layer thus produced were removed and the lower,egg-containing layer was diluted in a small amount of 1.2% saline and passed through a 325-mesh nylon net.The crude eggs preparation collected from above the net were layered over a dilution of Percoll with a density of 1.070 and centrifuged at 3 000 r/min for 20 min,and then the purified eggs in lower layer were collected.Furthermore,the purity of the separated eggs was detected under light microscope,the viability was identified by staining with acridine orange dye and circumoval precipitation test.The experiments had showed that about 1 gram eggs were recovered from a schistosoma-infected rabbit liver,the purity of the egg population was substantially improved,and the rate of live and viable eggs was much greater than more conventional filtration-centrifugation techniques.Therefore,the technique presented offers a simple and rapid method of obtaining highly purified preparations of Schistosoma japonicum eggs.The isolated eggs maintain their morphological,functional,and antigenic integrity.This method holds promise of being suitable for adaptation and expansion to a variety of experimental purposes.

关 键 词：日本血吸虫 虫卵 分离 纯化 活性 

分 类 号：R383.2[医药卫生—医学寄生虫学]