Zhang S, Wei M, Zhao H, Shi G
Shanghai Detecting Center for Toxoplasmosis, Shanghai Institute of Parasitic Disease Control and Rsesarch, Shanghai 200336.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1999;17(4):225-7.
To establish an immunosorbent agglutination assay (ISAGA) for detection of IgM antibodies against Toxoplasma gondii.
In the ISAGA, wells of microtiter plates were coated with anti-human IgM antibodies and sealed with 1% bovine serum albumin. After the test sera were added and incubated, the plates were washed, T. gondii tachyzoite antigen suspension was added, and incubated overnight at 37 degrees C. The ISAGA results were evaluated by comparing with those detected by Danish ISAGA and ELISA and those detected by slide enzyme immunoassay (S-EIA).
Forty-four sera from Danish pregnant women were tested by the IgM-ISAGA, 41(93.2%) were consistent with the Danish results. Sixty-seven sera from Danish and Shanghai pregnant women were detected by IgM-IgM-ISAGA and S-EIA, the total consistency rate was 92.5%. A significant correlation was found between the titers of the ISAGA and S-EIA, with a Pearson correlation coefficient of 0.589(P < 0.001). The titers of ISAGA were eighteen times higher than those of S-EIA. This method enables the detection of IgM antibodies as low as approximately 0.08 IU/ml.
The IgM-ISAGA is therefore sensitive, specific, easy to perform, and is useful for mass screening and diagnosing recent Toxoplasma infection or reactivation.
建立一种用于检测抗弓形虫IgM抗体的免疫吸附凝集试验(ISAGA)。
在ISAGA中,微量滴定板孔用抗人IgM抗体包被,并用1%牛血清白蛋白封闭。加入待测血清并孵育后,洗涤平板,加入弓形虫速殖子抗原悬液,于37℃孵育过夜。通过与丹麦ISAGA和ELISA检测结果以及玻片酶免疫测定(S-EIA)检测结果进行比较来评估ISAGA结果。
用IgM-ISAGA检测44份丹麦孕妇血清,41份(93.2%)与丹麦检测结果一致。用IgM-IgM-ISAGA和S-EIA检测67份丹麦和上海孕妇血清,总符合率为92.5%。ISAGA和S-EIA的滴度之间存在显著相关性,Pearson相关系数为0.589(P<0.001)。ISAGA的滴度比S-EIA高18倍。该方法能够检测低至约0.08 IU/ml的IgM抗体。
因此,IgM-ISAGA灵敏、特异、易于操作,可用于大规模筛查以及诊断近期弓形虫感染或再激活。