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.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
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)
关键词
日本血吸虫
虫卵
分离
纯化
活性
Schistosoma japonicum
eggs
isolation
purification
viability
