刁琳琪, 王伟, 徐建国, 廖兴广, 王芸. 胶体金标记免疫层析法检测大肠杆菌O157[J]. 疾病监测, 2007, 22(7): 438-440. DOI: 10.3784/j.issn.1003-9961.2007.7.438
引用本文: 刁琳琪, 王伟, 徐建国, 廖兴广, 王芸. 胶体金标记免疫层析法检测大肠杆菌O157[J]. 疾病监测, 2007, 22(7): 438-440. DOI: 10.3784/j.issn.1003-9961.2007.7.438
DIAO Lin-qi, WANG Wei, XU Jian-guo, LIAO Xing-guang , . et al.. Gold-immunochromatography assay for Escherichia coli O157[J]. Disease Surveillance, 2007, 22(7): 438-440. DOI: 10.3784/j.issn.1003-9961.2007.7.438
Citation: DIAO Lin-qi, WANG Wei, XU Jian-guo, LIAO Xing-guang , . et al.. Gold-immunochromatography assay for Escherichia coli O157[J]. Disease Surveillance, 2007, 22(7): 438-440. DOI: 10.3784/j.issn.1003-9961.2007.7.438

胶体金标记免疫层析法检测大肠杆菌O157

Gold-immunochromatography assay for Escherichia coli O157

  • 摘要: 目的 应用抗大肠杆菌O157(E.coli O157)单克隆抗体,制备一种简便快速的胶体金标记免疫层析(GICA)条用于检测标本中E.coli O157.方法 采用柠檬酸三钠还原法制备胶体金颗粒,标记抗E.coli O157单克隆抗体,以硝酸纤维膜作为抗E.coli O157单克隆抗体的包被载体,制成GICA检测条.标本中E.coli O157与检测条上金标记抗体(Au-Ab)结合后,利用硝酸纤维膜的层析作用移动,与膜上的固相抗体结合形成可见的红色条带.结果 GICA检测条灵敏度可达105 cfu/ml.用GICA检测了1750份不同食品和人畜粪便标本中E.coli O157,GICA检测条阳性份数43份,后经分离培养检出29株大肠杆菌O157;另14份标本经鉴定,其中12份标本可疑为弗劳地枸橼酸杆菌.结论 GICA条检测标本的E.coli O157,特异性较高,不需任何仪器设备,较简便快速,易于判断结果,故可对标本进行快速筛查,适合各级疾控中心和临床检验部门使用.

     

    Abstract: Objective The present study was conducted to establish a sensitive,rapid,and convenient gold immunochromatography assay (GICA) by using monoclonal anti-E.coli O157 for the detection of E.coli O157. Methods The trisodium citrate reduction method was used to prepare colloidal golds,which were coupled with monoclonal anti-E.coli O157. These antibodies were immobilized on nitrocellulose (N.C) membrane to prepare a GICA card device. The gold-antibodies (Au-Ab) conjugates reacted with E.coli 0157 in the samples and migrated up the N.C membrane to produce a optical red strip. Results The sensitivity of GICA was 105 cfu/ml. 1750 different food and stool samples were detected by GICA for E.coli 0157 and 43 samples were found to be GICA positive. E.coli O157 were isolated from 29 of the 43 positive samples with 12 C.freundii identified in the remaining 14 positive samples. Conclusion GICA is a specific,rapid and simple assay for the detection of E.coli O157,applicable to performing the tests in centers for disease control and prevention and clinical laboratories at various levels.

     

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