摘要
用胃蛋白酶消化旋毛虫感染鼠肌肉,获得肌肉期幼虫。37℃,用RPMI1640培养基培养幼虫,控制虫体死亡率低于5%。每24hr收集上清培养液,即为分泌排泄抗原(ESA)。共获10天次ESA(ESA_(1d~10d))。应用SDS-PAGE及氨银染色技术进行蛋白组份分析显示:各天次ESA的主要蛋白区带数及位置基本相同,分子量范围在96~14kd,主带3条,分别为48,53和58kD蛋白组份。应用Western-blot技术进行免疫识别分析显示:旋毛虫病人血清对各天次ESA的免疫识别结果相似,识别的抗原组份有7~8条区带,均有上述3条主带。正常人、蛔虫病人和囊虫病人血清均未能识别任何区带。肺吸虫病人血清能识别2~3条抗原区带,但其分子量位于24~14kD之间。血吸虫病人血清也能识别2~3条抗原区带,其分子量位于18~14kD之间。结果表明:在严格控制死亡虫体抗原污染条件下,较长期内培养肌肉期幼虫获得的各天次ESA,其成份和免疫识别效果基本无改变,各天次ESA中的46~58kD蛋白均具有良好的特异性。
Muscle larvae were obtained from infected mice by peptic digestion. The larvae were culturedin RPMI1640 medium at 37℃. The medium was changed every 24 hours for 10 days in order to remove thedead larvae Trichinella spiralis,and the rate of dead larvae must be no more than 5%。The collected super-natant,excretory-secretory antigen:ESA1d,ESA26,ESA46,ESA6d,ESA8d and ESA10d were analysed bySDS-PAGE with silver staining,Results showed that the stained bands of these kinds of ESA were similar innumber and site. The molecular weight of these banks were between 14~96 kD with three chief bands in 48,53 and 58 kD.ESA1d,ESA2d,ESA5d,ESA8d and ESA10d were analysed by Western-blot and the results showedthat the effect of immune recognition were similar.The sera from patients with trichinosis were recognizedwith 7~8 antigenic bands in chief bands of 46,48,50/53 and 58 kD. The sera from health persons and pa-tients with ascariasis or cysticerosis showed no any bands The sera from patients with paragonimiasis wererecognized with 2~3 antigenic bands at 14~24 kD,and those from patients with schistosomiasis with 2~3antigenic bands at 14~18 kD. These results indicate that the proteins of ESA between 46~58 kD were highlyspecific antigens.
出处
《地方病通报》
1996年第3期15-18,共4页
Endemic Diseases Bulletin
基金
卫生部基金
关键词
旋毛虫
肌肉期幼虫
分泌
排泄物
特异性抗原
分析
Trichinella spiralis
Muscle larvae
Excretory-secretory products
Specific antigens
Analysis