摘要
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、广州管圆线虫感染病例血清循环抗原（CAg），同时将12D5和2187单抗点样于固相的硝酸纤维膜上，以胶体金一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IgG1，2187为IgM，两株单抗同时识别广州管圆线虫成虫相对分子质量55×10^3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阋线虫的大鼠血清巾CAg检出率为100％（48／48），广州管圆线虫感染病例血清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。
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 21B7 McAbs could be prepared in advance. Methods Two McAbs （12D5 and 21B7） 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 ELISA. Either 12D5 or 21B7 McAbs was used as antibody and protein A was conjugated with colloid gold as the detection marker. A special pad for GICA was designed according to the reaction procedure, and CAg were detected by GICA in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively. Results 12D5 McAb was identified as IgG1 and 21B7 McAb was IgM. Results from Western blotting showed that two McAbs could be used to identified 55 KD protein of adult worms of A. cantonensis. The detection rates of CAg in the sera of infected rats was 100% （48/48） and the detection rates of CAg in the sera of angiostrongyliasis 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 normal sera did not react with 12D5 and 21B7 McAbs. Conclusion Results from sandwich ELISA and GICA with 12D5 and 21B7 McAbs showed high specificity and acting as detecting CAg of A. cantonensis in sera of infected animals and patients. We noticed that GICA with 12D5 and 21B7 was not only rapid and simple that without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A. cantonensis.
出处
《中华流行病学杂志》
CAS
CSCD
北大核心
2010年第1期79-82,共4页
Chinese Journal of Epidemiology
基金
广东省自然科学基金（8151802003000002）
关键词
广州管圆线虫
单克隆抗体
金标层析法
Angiostrongylus cantonensis
Monoclonal antibodies
Gold immunochromatography assay
