胶体金免疫层析法快速诊断鼠疫的实验研究  

Gold Immunochromatographic Assay for a Rapid Diagnosis for Yersinia pestis

作　　者：王鹏[1] 杜春红[1] 郭英[1] 尹家祥[1] 石丽媛[1] 陈平[1] 唐雪[1] 钟佑宏[1] 黄鹏[1] 吴明寿[1] 董兴齐[1] 于国林[1] 

机构地区：[1]云南省地方病防治所,云南大理671000

出　　处：《地方病通报》2006年第1期1-3,5,共4页Endemic Diseases Bulletin

基　　金：国家自然科学基金(30160078);云南省重大攻关项目(2002NG17)

摘　　要：目的建立一种快速、简易的检测鼠疫F1抗原的胶体金免疫层析法(G ICA)。方法采用直径为20nm胶体金颗粒,标记F1单克隆抗体,并将标记物以6μL/cm喷于玻璃纤维;将另1株F1单克隆抗体与羊抗鼠IgG以1μL/cm喷线固定于硝酸纤维素膜上,装配后以4mm/条切割制成免疫层析检测试条。分别用粗制F1抗原和EV菌对该试纸条的敏感性进行测定,同时用44株鼠疫近缘菌及昆明小鼠、黄胸鼠、褐家鼠、大足鼠的正常血清等标本进行特异性检测。结果本试纸条可在15m in之内完成实验,检测粗制F1抗原的浓度可达0.5ng/mL,最小检出EV菌菌量为10万个;本方法的特异性为100%。结论G ICA检测鼠疫F1抗原特异性强、灵敏度高、简便快速,无需特殊仪器设备,有广泛应用价值。Objective To develop a rapid, simple and self - performing gold immunoehromatographic assay （ GICA ） for the detection of F1 capsular antigen of Yersinia pestis. Methods We labeled monoelonal antibodies to the F1 antigen of Yersinia pestis with colloidal gold （ diameter 20nm ） and sprayed them to fiberglas by 6μL/cm, sprayed another F1 monoclonal antibodies and goat- anti -mouse IgG to nitrocellose strip, and assembled them and incised into lateral test strips. Its sensitivity and specificity were determined with crude F1 antigen, EV strain, 44 strains of rela：ive bacteria and the normal serum of Kunming mice, R.flavipectus, R. rtorvegieus and R. nitidus, and so on. Results The lateral test strips detected concentrations of F1 antigen as low as 0.5 ng/mL in 15 min, and its ：pecificity were 100% tested with the above specimen. Conclusions GICA is a sensitive, specific, simple and rapid assay for detecting F1 capsular antigen of Yersinia pestis.

关 键 词：鼠疫 胶体金 免疫层析 F1抗原 

分 类 号：R516.8[医药卫生—内科学] R446[医药卫生—临床医学]