Enhancing sensitivity in colorimetric immunoassay by using secondary antibody-gold nanoparticle conjugate

2013 13th IEEE International Conference on Nanotechnology (IEEE-NANO 2013) Pub Date : 2013-08-01 DOI:10.1109/NANO.2013.6720846

W. Maneeprakorn, C. Apiwat, T. Dharakul A

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引用次数: 2

Abstract

A lateral flow colorimetric immunoassay (LFA) based on gold nanoparticles (AuNP) for detecting the antibody-antigen interaction and gold enhancement for amplifying the specific binding signal has been developed. Influenza A was used as the model study to determine the performance of the system. Nucleoprotein (NP) of the influaza A virus was selected as the targeting molecule. Two different sizes of AuNP conjugated to antibody against NP were prepared and applied to the assay. The 1st conjugate was AuNP immobilized with an anti-NP antibody and secondary-immobilized with biotinylated bovine serum albumin (B-BSA). The 2nd conjugate was AuNP immobilized with an anti-biotin antibody. The detection sensitivity of the enhanced sensitivity LFA increased about 3-folds and 8-folds compared with conventional LFA and commercial influenza A LFA, respectively. The signal intensity of the test spot/line at the capture test zone was increased dramatically. With this method, nucleoprotein of influenza A virus can be detected within 15 minutes and was highly reproducible. The results indicated that the technique can facilitate the more sensitive LFA test for diagnosis of many diseases.

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利用二抗-金纳米颗粒偶联物提高比色免疫分析的灵敏度

建立了一种基于金纳米颗粒(AuNP)的横向流动比色免疫分析法(LFA)，用于检测抗体-抗原相互作用和放大特异性结合信号的金增强。以甲型流感为模型研究确定系统的性能。选择甲型流感病毒核蛋白(NP)作为靶分子。制备了两种不同大小的与NP抗体结合的AuNP，并将其用于检测。第一个偶联物是用抗np抗体固定AuNP，用生物素化牛血清白蛋白(B-BSA)二次固定。第二个偶联物是用抗生物素抗体固定的AuNP。与传统LFA和商用流感A LFA相比，增强敏感性LFA的检测灵敏度分别提高了3倍和8倍。捕获试验区测试点/线的信号强度显著增加。该方法可在15分钟内检测出甲型流感病毒核蛋白，重现性好。结果表明，该技术可使LFA检测对多种疾病的诊断更为灵敏。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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2013 13th IEEE International Conference on Nanotechnology (IEEE-NANO 2013)

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