Jump to main content
Jump to site search

Issue 23, 2014
Previous Article Next Article

Development of a highly sensitive real-time immuno-PCR for the measurement of chloramphenicol in milk based on magnetic bead capturing

Author affiliations

Abstract

Chloramphenicol (CAP) is a forbidden antibiotic that enters the food chain by illegal use in food-producing animals, potentially causing aplastic anaemia in humans. Immuno-PCR has the potential to address the need of meeting strict limits for this antibiotic by detecting trace levels of CAP present in animal-derived foods. A real-time immuno-quantitative PCR (RT-IPCR) assay for the quantification of CAP based on the simple and quick immunomagnetic bead recovery of CAP in milk was developed. The immunomagnetic beads were obtained by linking the reporter DNA to anti-CAP monoclonal antibody with N-succinimidyl S-acetylthioacetate (SATA) and succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) and immobilising the anti-CAP monoclonal antibody/DNA-label conjugate on the magnetic beads with 1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (SNHS). The RT-IPCR assay allows the sensitive detection and quantification of CAP in the concentration range of 0.001–0.11 μg L−1 (R2 = 0.9986) with a 50% inhibition concentration (IC50) value of 0.008 μg L−1. The RT-IPCR approach discussed here is presented as a model system that could be easily adapted for small molecule detection in a variety of foods using simple immunomagnetic bead recovery.

Graphical abstract: Development of a highly sensitive real-time immuno-PCR for the measurement of chloramphenicol in milk based on magnetic bead capturing

Back to tab navigation

Supplementary files

Publication details

The article was received on 15 Sep 2014, accepted on 08 Oct 2014 and first published on 08 Oct 2014


Article type: Paper
DOI: 10.1039/C4AY02158D
Author version
available:
Download author version (PDF)
Citation: Anal. Methods, 2014,6, 9340-9347
  •   Request permissions

    Development of a highly sensitive real-time immuno-PCR for the measurement of chloramphenicol in milk based on magnetic bead capturing

    X. Tao, Z. He, X. Cao, J. Shen and H. Li, Anal. Methods, 2014, 6, 9340
    DOI: 10.1039/C4AY02158D

Search articles by author

Spotlight

Advertisements