Journal
MICROCHIMICA ACTA
Volume 180, Issue 3-4, Pages 303-310Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-012-0934-y
Keywords
Porcine pseudorabies virus; Immunoassay; Fluorescence signal amplification; Cation-exchange; CdSe nanocrystals
Categories
Funding
- National Natural Science Foundation of China [20975042, 21175051]
- Fundamental Research Funds for the Central Universities [2010PY009, 2011PY139]
- Natural Science Foundation of Hubei Province Innovation Team [2011CDA115]
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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.
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