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Magnetic Bead-Based Sandwich Immunoassay for Viral Pathogen Detection by Employing Gold Nanoparticle as Carrier

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Abstract

Enzyme-linked immunosorbent assay (ELISA) provides a convenient way for the detection of viral pathogens. However, conventional ELISA performed on mirowell plates suffers from poor sensitivity, laborious coating and complicated blocking procedures. Herein, we designed a sensitive colorimetric immunoassay by taking advantages of the enrichment and isolation ability of magnetic beads (MBs) and the high loading capacity of gold nanoparticles (AuNPs) for detecting respiratory syncytial virus (RSV) as a pathogen model. RSV was selectively captured and preconcentrated from samples with antibodies functionalized MBs, followed by binding with antibodies labeled AuNPs, which carrying a large amount of alkaline phosphatase (ALP) molecules for colorimetric signal amplification by catalyzing the dephosphorylation of non-colored pNPP to generate colored product pNP. After optimizing the experimental conditions based on the principle of low nonspecific signal, low cost, and high sensitivity, the analytical sensitivity of the developed immunoassay can be improved to 0.27 pg/mL, which is over sevenfold higher than that of commercially available RSV ELISA kits (2 pg/mL). In addition, the total assay time was less than 2.5 h without any pretreatment, which is much more rapid than other reported assays. Therefore, the proposed immunoassay holds great promise for the fabrication of rapid, sensitive, and economic method for the viral pathogen detection.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC, No. 21535006), the National Basic Research Program of China (973 Program, No. 2011CB933600), and the Doctoral Scientific Research Foundation (SWU116058).

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Authors and Affiliations

  1. Key Laboratory on Luminescence and Real-Time Analytical Chemistry (Ministry of Education), College of Pharmaceutical Science, Southwest University, Chongqing, 400716, China

    Lei Zhan, Wen Bi Wu, Chun Mei Li & Cheng Zhi Huang

  2. Chongqing Key Laboratory of Biomedical Analysis (Chongqing Science & Technology Commission), College of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, China

    Cheng Zhi Huang

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  1. Lei Zhan
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  2. Wen Bi Wu
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  3. Chun Mei Li
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Correspondence to Chun Mei Li or Cheng Zhi Huang.

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Zhan, L., Wu, W.B., Li, C.M. et al. Magnetic Bead-Based Sandwich Immunoassay for Viral Pathogen Detection by Employing Gold Nanoparticle as Carrier. J. Anal. Test. 1, 298–305 (2017). https://doi.org/10.1007/s41664-017-0040-6

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  • DOI: https://doi.org/10.1007/s41664-017-0040-6

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