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Development of indirect competitive ELISA using egg yolk-derived immunoglobulin (IgY) for the detection of Gentamicin residues.

Posted by on in 2016
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He J1,2Hu J1Thirumalai D2Schade R2Du E2Zhang X1,2 2016. J Environ Sci Health B. 51(1):8-13. doi: 10.1080/03601234.2015.1080479. Epub 2015 Oct 29.

  • 1a College of Animal Science, Tarim University , Alaer, Aksu Xinjiang , China.
  • 2b College of Veterinary Medicine, Northwest A&F University , Yangling, Shaanxi , China.


Gentamicin (Gent) is an aminoglycoside antibiotic being used in livestock sector. Gent residues could cause some genetic disorders by nonsense mutations. This study aimed to develop IgY-based ELISA for the detection of Gent in animal products. Gent was conjugated with Bovine serum albumin (BSA) by carbodiimide method for further immunization in the laying chickens. PEG-6000 extraction method was employed to extract IgYfrom the egg yolk. The titer of anti-Gent-IgY attained the peak of 1:256,000 after the 5(th) booster immunization. Checkerboard titration confirmed that, anti-Gent IgY in 1:2,000 dilution could give an Optical Density (OD) 1.0 at 2 µg mL(-1) of Gent-OVA coating concentration. IgY-based indirect competitive ELISA (Ic-ELISA) showed that, the IC50 value of anti-Gent IgY was 2.69 ng mL(-1) and regression curve equation was y = -16.27x + 56.97 (R(2) = 0.95, n = 3), confirming that, the detection limit (LOD, IC10 value) was 0.01 ng mL(-1). Recoveries from fresh milk, pork and chicken samples were ranged from 69.82% to 94.32%, with relative standard deviation lower than 10.88%. Our results suggested that generated anti-Gent IgYantibodies can be used in routine screening analysis of Gent residues in food samples.


Drug residues; ELISA; gentamycin; yolk immunoglobulin (IgY)

[PubMed - in process]
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