摘要
为减少弓形虫假包囊和宿主细胞对试验的干扰,对弓形虫的速殖子进行了分离纯化。将弓形虫RH株虫体复苏后,接种于小鼠腹腔进行培养,72h后收集腹液,腹液经27G注射器针头抽吸后进行不同组合的非连续密度梯度(1.031×106 mg/L、1.056×106 mg/L、1.077×106 mg/L、1.123×106 mg/L Percoll)离心纯化。通过显微镜观察以及细胞计数等方法比较各组合的纯化效果。结果显示,经注射器针头帮助假包囊破裂后,使用非连续密度梯度离心,以密度为1.056×106 mg/L、1.077×106 mg/L Percoll不连续梯度3 000r/min离心30min,可以将速殖子和假包囊进行较高纯度的分离纯化。本研究为纯化高纯度的弓形虫速殖子提供了一种方法。
In order to avoid the interferences from the host cells and pseudocysts during some experiments,the method to separate and purify the tachyzoites of Toxoplasma gondii from host cells was explored.RH strain of T.gondii was propagated by intra-peritoneal injection in mice,and the tachyzoites were harvested from peritoneal cavity after 72 hours post infection.The peritoneal fluid containing tachyzoites,pseudocysts and host cells was passed through a 27-gauge needle before centrifugation on a discontinuous Percoll density-gradience which was presented by different density combinations(1.031×106 mg/L,1.056×106 mg/L,1.077×106 mg/L,1.123×106 mg/L).Results suggested that the method of discontinuous density gradient centrifugation could separate the tachyzoites from pseudocysts effectively.This study provided a method for the separation of the tachyzoites of T.gondii with high purity.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2012年第5期467-472,共6页
Chinese Veterinary Science
基金
"十一五"传染病重大专项(2008ZX10004-011)
关键词
刚地弓形虫
速殖子
假包囊
分离纯化
Toxoplasma gondii
tachyzoite
pseudocyst
separation and purification