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Sensitive immunoassay for porcine pseudorabies antibody based on fluorescence signal amplification induced by cation exchange in CdSe nanocrystals

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Abstract

We report on a novel immunoassay for porcine pseudorabies virus (PRV) antibody that is based on fluorescence signal amplification induced by silver(I) ion exchange in CdSe nanocrystals. An antigen-antibody-secondary antibody sandwich structure was first formed from PRV, PRV antibody, and CdSe-labeled rabbit anti-pig antibody. Then, the Cd(II) ions in the CdSe labels were released by a cation exchange reaction with Ag(I). Released Cd(II) was finally quantified using the sensitive fluorescent probe Rhodamine 5 N. Due to this signal amplification, the sensitivity and linear range of the immunoassay were largely improved (compared to the traditional ELISA) in having a limit of detection as low as 1.2 ng mL−1 of PRV antibody and a linear range from 2.44 to 312 ng mL−1. The successful determination of PRV antibody in pig serum samples is proof for the utility of the method.

A simple, rapid and sensitive method for the detection of PRV antibody through the fluorescence signal amplification caused by cation-exchange in CdSe NCs was reported. The CdSe NCs labeled rabbit anti-pig IgG was used to capture the PRV antibody. After the immunoreaction, the Cd2+ in the CdSe labels was completely replaced by the cation-exchange reaction with Ag+. Then Cd2+sensitive fluorescence indicator Rhod-5 N was added to bind with Cd2+ and caused the fluorescence signal enhance substantially. Thus a novel method for rapid and sensitive detection of porcine pseudorabies based on the fluorescence signal amplification was developed.

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Acknowledgements

The authors gratefully acknowledge the support for this research from National Natural Science Foundation of China (20975042, 21175051), the Fundamental Research Funds for the Central Universities (2010PY009, 2011PY139) and the Natural Science Foundation of Hubei Province Innovation Team (2011CDA115).

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Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Science, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    XuePu Li, Kun Chen, Liang Huang, DongLian Lu, JianGong Liang & HeYou Han

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  1. XuePu Li
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  2. Kun Chen
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  3. Liang Huang
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  4. DongLian Lu
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  5. JianGong Liang
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  6. HeYou Han
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Correspondence to HeYou Han.

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Li, X., Chen, K., Huang, L. et al. Sensitive immunoassay for porcine pseudorabies antibody based on fluorescence signal amplification induced by cation exchange in CdSe nanocrystals. Microchim Acta 180, 303–310 (2013). https://doi.org/10.1007/s00604-012-0934-y

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  • DOI: https://doi.org/10.1007/s00604-012-0934-y

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