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

被引：28

作者：

He, Jinxin ^{[1
,2
]}

Hu, Jianjun ^{[1
]}

Thirumalai, Diraviyam ^{[2
]}

Schade, Ruediger ^{[2
]}

Du, Enqi ^{[2
]}

Zhang, Xiaoying ^{[1
,2
]}

机构：

[1] Tarim Univ, Coll Anim Sci, Alaer, Aksu Xinjiang, Peoples R China

[2] Northwest A&F Univ, Coll Vet Med, Yangling 712100, Shaanxi, Peoples R China

来源：

JOURNAL OF ENVIRONMENTAL SCIENCE AND HEALTH PART B-PESTICIDES FOOD CONTAMINANTS AND AGRICULTURAL WASTES | 2016年 / 51卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Drug residues; ELISA; gentamycin; yolk immunoglobulin (IgY); ANTIBIOTICS; ASSAY; MILK; AMINOGLYCOSIDES; CHROMATOGRAPHY; ANTIBODIES;

D O I：

10.1080/03601234.2015.1080479

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

摘要：

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 IgY from 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 mu g mL(-1) of Gent-OVA coating concentration. IgY-based indirect competitive ELISA (Ic-ELISA) showed that, the IC50 value of anti-Gent IgY was 2.69ng 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 0.01ng 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 IgY antibodies can be used in routine screening analysis of Gent residues in food samples.

引用

页码：8 / 13

页数：6

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