Development and application of one-step ELISA for the detection of neomycin in milk

被引：38

作者：

Xu, Naifeng ^{[1
,2
]}

Qu, Changlong ^{[1
,2
]}

Ma, Wei ^{[1
,2
]}

Xu, Liguang ^{[1
,2
]}

Liu, Liqang ^{[1
,2
]}

Kuang, Hua ^{[1
,2
]}

Xu, Chuanlai ^{[1
,2
]}

机构：

[1] Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China

[2] Jiangnan Univ, Sch Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China

来源：

FOOD AND AGRICULTURAL IMMUNOLOGY | 2011年 / 22卷 / 03期

关键词：

neomycin; polyclonal antibody; ELISA; milk; LINKED-IMMUNOSORBENT-ASSAY; LIQUID-CHROMATOGRAPHY; RESIDUE; GENTAMICIN; HEXOESTROL; AMINOGLYCOSIDES; PYRETHROIDS;

D O I：

10.1080/09540105.2011.569882

中图分类号：

O69 [应用化学];

学科分类号：

070301 [无机化学];

摘要：

A sensitive enzyme-linked immunosorbent assay (ELISA) for the detection of neomycin (NEO) in milk was developed. Two conjugates were synthesized as the immunogen and coating antigen. Four BALB/c female mice were immunised and the antisera were screened by indirect competitive ELISA (icELISA). The icELISA was further optimised using different coating methods, pH values, ionic strengths of assay buffer and reaction times. After optimised, the indirect competitive ELISA (icELISA) could be finished in 85min with an IC50 value of 0.74 +/- 0.05 ng/mL. One-step indirect competitive ELISA (icELISA) exhibited similar sensitivity with an IC50 value of 0.73 +/- 0.03 ng/mL, which was sensitive enough for NEO detection and could be finished in 55 min. No cross-reactivity of the antibody was observed with other aminoglycosides based on icELISA, indicating that the antibody is highly specific for neomycin. Milk samples were further analyzed with recovery ranging from 85 to 110% at spiked level of 0.25-10 ng/mL, and the detection limit was 0.08 ng/mL.

引用

页码：259 / 269

页数：11

共 24 条

[1]

[Anonymous], EMEAMRL81602FINAL

[2]

Comparative study on the analytical performance of three waveforms for the determination of several aminoglycoside antibiotics with high performance liquid chromatography using amperometric detection [J].