| 黄达娜,陈木新,耿艺介,李晓恒,高世同,张仁利.应用12D5和2187单克隆抗体检测广州管圆线虫循环抗原的研究[J].Chinese journal of Epidemiology,2010,31(1):79-82 |
| 应用12D5和2187单克隆抗体检测广州管圆线虫循环抗原的研究 |
| Detection of circulating antigen of Angiostrongylus cantonensis by 12D5 and 2187 monoclonal antibodies |
| Received:May 12, 2009 |
| DOI： |
| KeyWord: 广州管圆线虫 单克隆抗体 金标层析法 |
| English Key Word: Angiostrongylus cantonensis Monoclonal antibodies Gold inununochromatography assay |
| FundProject:广东省自然科学基金(8151802003000002) |
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| Abstract: |
| 目的研制快速诊断广州管圆线虫感染的金标层析检测试剂。方法双抗体(12D5和2187)夹心ELISA检测实验感染广州管圆线虫的大鼠、广州管网线虫感染病例血清循环抗原(CAg)，同时将12D5和2187单抗点样于同相的硝酸纤维膜上，以胶体金-proteinA为标记物制备金标免疫层析试剂快速检测实验感染广州管圆线虫的大鼠、广州管圆线虫感染病例血清循环抗原。结果经鉴定单抗12D5为IgGl，2187为IgM，两株单抗同时识别广州管围线虫成虫相对分子质量55×103的蛋白，12D5和2187双抗体夹心ELISA和金标层析法对实验感染广州管网线虫的大鼠血清中CAg检出率为100％(48/48)，广州管囤线虫感染病例血清CAg检出率为100％(32/32)，与日本血吸虫、肝吸虫、肺吸虫、旋毛虫、蛔虫、包虫病例血清无交叉反应，与健康人血清无反应。结论12D5、2187双抗体夹心ELISA和金标层析法对感染广州管圆线虫人和动物血清中CAg检测的特异性强，金标层析法操作简便、快速，结果判读容易，不需特殊设备，且敏感性高，能够确定现症感染。 |
| English Abstract: |
| Objective To detect the rate ofAngiostrongylus cantonensis infection and to study the effects of treatment so as to prepare monoclonal antibodies(McAbs)，and gold immunochromatography assay(GICA)with 12D5 and 2187 McAbs could be prepared in advance.Methods Two McAbs(12D5 and 2187)were applied to detect the circulating antigen(CAg)in the sera of rats infected with A.cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELlSA.Either 12D5 or 2lB7 McAbs was used as antibody and protein AWas conjugated with colloid gold as the detection marker.A special pad for GICA Was designed according to the reaction procedure。and CAg were detected bv GICA in the sera of rats infected with A eantonemis and angiostrongyliasis patients respectively.Results 12D5 McAb Was identified as IgGl and 2187 McAb Was IgM.Results from Westem blotting showed that two McAbs could be used to identified 55 KD protein of adult worms of A.eantonemis.The detection rates of CAg in the sera of infected rats was 100％(48/48)and the detection rates ofCAg in the sera ofangiostrongTliasis patients Was 100％ (32/32).NO cross-reaction to sera of patients with other infection of parasites.such as clonochiasis。 fasiolopsiasis，ancylostomiasis，ancylostomiasis.anisakiasis as well as schsitosomiasis wee seen and norrnal sera did not react with 1 2D5 and 2 1 B7 McAbs.Conclusion Results from sandwich ELISA and GICA with 1 2D5 and 2 1 B7 McAbs showed high specificity and acting as detecting CAg of A eantoneasis in sera of infected animals and patients.We noticed that GICA witll l 2D5 and 2 l B7 Was not only rapid and simple that without requirement of special instrument。but also rather sensitive and specific for the detection ofcurrent infeetion with A.cantonensis. |
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