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Gold nanoparticles based lateral flow immunoassay with largely amplified sensitivity for rapid melamine screening

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Abstract

The sensitivity of current lateral flow strip (LFS) based assays is a technical bottleneck. The authors describe a method for signal amplification to give a 10- to 25-fold improvement of the detection limit without compromising the assay time and operational mode. The assays includes the following steps: (a) immobilization of melamine antibody on gold nanoparticles (AuNPs); (b) immobilization of BSA antibody on other AuNPs; (c) coating a nitrocellulose (NC) membrane with coating antigen and secondary antibody against the melamine antibody as the test line and control line, respectively; (d) introducing an enhancement pad for signal amplification by immuno-recognition of the blocking protein on the first conjugation pad by the antibody on the enhancement pad. This results in dual labeling of AuNPs on the test line and enhances the optical intensity of test line at the same time of detection. The signal amplification makes the optical intensity change of the test line distinguishable even at low concentrations. The assay has a detection limit of 1.4 ppb (at an S/N ratio of 3). It was applied to the determination of melamine in milk, and the results obtained with this LFS are in good agreement with those obtained with instrumental methods. This demonstrates the practical applicability of the amplified LFS to real food samples. In our perception, this approach has a wide scope because it paves the way to the systematic amplification of the sensitivity of lateral flow strips.

Schematic image of signal amplified lateral flow strip (NC: nitrocellulose). One-step signal amplification strategy was adopted into traditional lateral flow strip to improve the sensing performance with the detection limit of 1.4 ppb for melamine, which is an about 25-fold improvement.

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Acknowledgment

This work is financially supported by the NSFC grant of 21475030, 31301460, the Science and Technology Research Project of Anhui Province 15czz03109, National 10000 Talents-Youth Top-notch Talent Program, the National and Zhejiang Public Benefit Research Project (201313010, 2014C32051) and the Jiangsu Science and Technology Support Program of BE201373, 2012780.

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Authors and Affiliations

  1. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei, 230009, China

    Youhao Zhong, Li Yao, Diping Zhao, Lei Zheng, Yingwang Ye & Wei Chen

  2. College of Food Science and Engineering/Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing University of Finance and Economics, Nanjing, 210023, China

    Yinji Chen

  3. Department of Chemistry & Biochemistry, North Dakota State University, Fargo, 58108, USA

    Guodong Liu

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  1. Youhao Zhong
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  2. Yinji Chen
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  3. Li Yao
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  4. Diping Zhao
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  5. Lei Zheng
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  6. Guodong Liu
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  7. Yingwang Ye
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  8. Wei Chen
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Correspondence to Yingwang Ye or Wei Chen.

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The author(s) declare that they have no competing interests.

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Youhao Zhong and Yinji Chen contributed equally to this manuscript.

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Zhong, Y., Chen, Y., Yao, L. et al. Gold nanoparticles based lateral flow immunoassay with largely amplified sensitivity for rapid melamine screening. Microchim Acta 183, 1989–1994 (2016). https://doi.org/10.1007/s00604-016-1812-9

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  • DOI: https://doi.org/10.1007/s00604-016-1812-9

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