胶体金免疫层析法与双单克隆抗体夹心ELISA检测鼠疫F1抗原的比较  

Comparison of Detecting F1 Antigen of Yersinia pestis Between Gold-immunochromatography Assay and Double Monoclonal Antibody Sandwich Enzyme-linked Immunosorbent Assay

作　　者：刘合智[1] 王海峰[1] 史献明[1] 杨晓燕[1] 周松[1] 白雪薇[1] 胡乐乐[1] 郑楠[1] 张懿晖[1] 董国润[1] 李玉贵[1] 

机构地区：[1]河北省鼠疫防治所,河北张家口075000

出　　处：《地方病通报》2010年第1期7-11,共5页Endemic Diseases Bulletin

基　　金：河北省卫生厅医学科学研究重点项目(2005080);河北省医学适用技术跟踪项目(GL200635)

摘　　要：目的比较胶体金免疫层析法(GICA)和双单克隆抗体夹心酶联免疫吸附试验(DMcAbS-ELISA)检测鼠疫F1抗原的敏感性和特异性。方法用GICA试纸条和DMcAbS-ELISA平行检测308份强毒鼠疫耶尔森菌感染鼠脏器标本和327份对照鼠标本,用RIHA微量法同时检测作为参照。结果327份对照鼠鼠疫细菌学检验及GICA,DMcAbS-ELISA和RIHA的F1抗原检测均为阴性,GICA,DMcAbS-ELISA和RIHA在108cfu/mL水平上未发现与近缘假结核耶尔森菌有交叉反应;308只感染鼠样本细菌培养阳性284只,24只样本未分离到鼠疫菌;F1抗原检测GICA阳性287份,DM-cAbS-ELISA阳性280份,RIHA阳性276份,GICA阳性检出率最高93.19%,与DMcAbS-ELISA和RIHA比较,差异均有统计学意义(χ2=5.14,9.09;P=0.016,0.001);GICA与DMcAbS-ELISA符合率97.73%(Kappa=0.845);GICA和DMcAbS-ELISA与RIHA符合率分别是96.43%和98.70%(Kappa=0.774,0.926);284份细菌培养阳性鼠标本F1抗原检测GICA阳性率是98.59%,高于DMcAbS-ELISA和RIHA,差异有统计学意义(χ2=4.17,5.14;P=0.031,0.016);24份细菌培养阴性实验鼠标本F1抗原检测:GICA检出7份阳性,阳性率29.17%,DMcAbS-ELISA检出6份阳性,阳性率25%,RIHA检出3份阳性,阳性率12.5%,GICA高于DMcAbS-ELISA和RIHA,但差异无统计学意义(χ2=2.25,0;P=0.125,1.000)。结论GICA检测鼠疫F1抗原敏感特异,快速简便,优于DMcAbS-ELISA和RIHA,是鼠疫快速诊断中有应用价值的检测技术。Objective To compare sensitivity and specificity of gold-immunochromatography assay（GICA） and double monoclonal antibody sandwich enzyme-linked immunosorbent assay（DMcAbS-ELISA） for detecting F1 antigen of Yersinia pestis.Methods Viscera organ（liver and spleen） specimens of 308 mice with virulent Yersinia pestis infection and 327 control samples of rats were detected with GICA dipstick and DMcAbS-ELISA.Meanwhile,micro-method of reverse indirect hemagglutination assay（RIHA） was carried out for parallel testing.Results Plague bacteriological tests of 327 control rats,F1 antigen detection with GICA,DMcAbS-ELISA and RIHA were negative.No cross-reaction with relatives of Yersinia pseudotuberculosis at 108 cfu/mL level was found in GICA,DMcAbS-ELISA and RIHA.Out of the 308 infected mice,284 were positive of plague bacterial culture,24 samples without isolation of Yersinia pestis.There were 287 of positive by GICA,280 of positive by DMcAbS-ELISA,276 of positive by RIHA.Positive detection rate of GICA was the highest of 93.19%,with statistically significant differences when compared with DMcAbS-ELISA and RIHA（χ^2=5.14,9.09;P=0.016,0.001）.Coincidence was 97.73% between GICA and DMcAbS-ELISA（Kappa=0.845）,and 96.43% and 98.70% between GICA,DMcAbS-ELISA and RIHA,respectively（Kappa = 0.774,0.926）.In 284 samples with positive bacterial culture,detection rate of F1 antigen was 98.59% by GICA,higher than that by DMcAbS-ELISA and RIHA,with statistically significant difference（χ^2=4.17,5.14;P=0.031,0.016）.Out of 24 specimens with negative bacterial culture for F1 antigen,there were 7 of positive detected by GICA with the positive rate of 29.17%,6 of positive by DMcAbS-ELISA with the positive rate of 25%,3 of positive by RIHA with the positive rate of 12.5%.The positive rate of GICA was higher than that of DMcAbS-ELISA and RIHA,but no statistically significant difference was found（χ^2 = 2.25,0;P=0.125,1.000）.Conclusions GICA is sensitive and specific,fast and simple in detection for F1 antigen

关 键 词：胶体金免疫层析试验 酶联免疫吸附试验 耶尔森氏菌 鼠疫 F1抗原 

分 类 号：R378.61[医药卫生—病原生物学] R446.6[医药卫生—基础医学]