日本血吸虫未成熟虫卵的制备

Preparation of Immature Eggs of Schistosoma japonicum

从感染兔肝组织中分离、纯化具有生物活性的日本血吸虫虫卵,是分子生物学研究的基本需要。为改进日本血吸虫(Schistosoma japonicum)未成熟虫卵的制备方法,将感染血吸虫尾蚴30d的家兔剖杀后,于无菌条件下摘取肝脏,经匀浆、分级过滤、中速低温离心沉淀、反复洗涤,再将粗虫卵置于0.25%胰蛋白酶消化液中消化4h,当混杂组织被消化成单个细胞后,用PBS(pH 7.2)稀释,以360目分样筛过筛,截留未成熟虫卵,获得大量高纯度、具有活性的虫卵,毛蚴孵出率可达80%以上。此法经济简便、快速易行、效率高,适于大批量操作。

It is essential to isolate and purify Schistosoma japonicum eggs with biological activity from the postinfective rabbit liver tissue for the study of molecular biology.To improve the method of preparing a large amount of pure alive Schistosoma japonicum eggs,the rabbit was sacrificed after being infected with 4500~5000 cercariae about 30 days and its liver was taken aseptically and homogenized.By the consecutive method of homogenization,fractional fictration,centrifugation,respective wash,the eggs and leftover were then digested with 0.25% trypsin for 4 hours,diluted with PBS(pH7.2) and sieved over 360 meshes per inch to keep the immature eggs.And highly purified eggs biological activity could be obtained with more than 80% incubative rate of miracidia.The results indicate that the method is more economical,convenient and effective as compared with previous ones,especially more suitable for isolation purification in a large scale.