双抗体夹心免疫-PCR检测弓形虫循环抗原的实验研究  

Study on the detection of Toxoplasma circulating antigens with double antibodies immuno-PCR

作　　者：华海涌[1] 周永华[1] 薛忠权[1] 

机构地区：[1]江苏省寄生虫病防治研究所,江苏无锡214064

出　　处：《中国现代医药杂志》2005年第6期1-2,共2页Modern Medicine Journal of China

基　　金：江苏省卫生厅资助课题(X200337)

摘　　要：目的探索弓形虫病的病原学检测方法。方法用游离的亲合素将生物素化的二抗和生物素化的腺病毒六邻体基因重组质粒DNA偶联起来,通过PCR扩增标记的DNA检测弓形虫循环抗原,建立了免疫-PCR检测方法;用该方法和ELISA法平行检测感染鼠的血清,对比研究两者敏感性及循环抗原的检出时间。结果免疫-PCR法检测弓形虫循环抗原阳性血清的最大稀释度为1∶2560,ELISA法为1∶10;免疫-PCR法在感染后第2天检测到循环抗原,ELISA法在第4天检测到循环抗原。结论免疫-PCR是一种特异性高、敏感性强的弓形虫循环抗原的检测方法,可作为弓形虫病病原学的诊断方法。Objective To explore an diagnosis assay for aetiology of Toxoplasmosis. Methods Free streptavidin was used to attach the biotinylated second antibody to the Bio-AdAT DNA,through amplication of the DNA lable, Toxoplasma circulating antigens was detected, thus Immuno-PCR assay was established; Detecting the serum of infected mice using Immuno- PCR and ELISA parallely to compare the sensitivity and the time of circulating antigens occuring. Results The detection limit of the Immuno-PCR was at about 1：2 560 dilution and it was at 1 ： 10 dilution of ELISA; Circulating antigens of Toxoplasma could be find as early as the second clay after infection using Immuno-PCR, but it was detected on the fouth clay by ELISA. Conclnsion Immuno-PCR could be used as a diagnosis method for aetiology because of its specificity and sensitivity for detecting the circulating antigens of Toxoplasma.

关 键 词：免疫-PCR 弓形虫 循环抗原 

分 类 号：R392[医药卫生—免疫学]