Dot-ELISA检测弓形虫循环抗原的方法学研究  

STUDY ON METHOLOGY OF DOT-ELISA FOR DETECTING CIRCULATING ANTIGENS OF TOXOPLASMA

作　　者：刘佩梅 杨树森[1] 杨秀珍[1] 朱静和[1] 

机构地区：[1]天津医学院寄生虫学教研室

出　　处：《中国寄生虫病防治杂志》1990年第3期220-223,共4页Chinese Journal of Parasitic Disease Control

基　　金：国家自然科学基金课题(3880714)

摘　　要：本文对 Dot-ELISA 检测弓形虫循环抗原(CAg)的方法学进行了研究。结果显示,其最佳试验条件为:纯化的特异抗体1;100稀释,与被检样本在37℃孵育2h;第二抗体-HRP 1:2000稀释,置室温30min;DAB 量5mg/10ml、H_2O_22μl。在该条件下,检测 TBS、正常的人血清、小鼠血清、猪血清、人尿和脑脊液内弓形虫抗原量,最低检出阈值依次是3.84pg/ml、96pg/ml、0.48ng/ml、0.48ng/ml、12ng/ml、12ng/ml,并呈良好的重复性。以5×10~3形虫 RH 株速殖子感染小鼠,d_3呈弱阳性,d_4明显阳性。用该条件检测人恶性疟原虫、旋毛虫和猪囊虫抗原,未呈现交叉反应。The antigens of Toxoplasma gondii were detected by dot-ELISA.Its optimum was as follows:specific antibodies at 1:100,incubation time about 2h at 37℃,HRP- conjugate at 1:2000 and incubating for 30 min at room temperature,TBS at pH7.4 and 3% gelatin-TBS blccking about 1h at room temperature.The sensitivity of detecting antigens of T.gondii was 3.84pg/ml in TBS,96pg/ml in normal human serum,0.48ng/ ml in normal mouse serum,0.48ng/ml in normal pig serum,12ng/ml in normal human urine,and 12ng/ml in normal human cerebrospinal fluid.There was a better reproduci- bility and no crcssreacticn among Plasmodium falciparum,Trichinella spiralis,Cysti- cercus cellulosae and T.gondii. when mice were infected with 5×10~3 tachyzoites of T.gondii RH strain,the anti- genemia was demonstrated by dot-FLISA at 3 rd day after infection.The reaction at 4th day became distinct pcsitive.

关 键 词：弓形虫病 循环抗原 DOT-ELISA 

分 类 号：R531.8[医药卫生—内科学]