Development of a sensitive indirect competitive enzyme-linked immunosorbent assay based on the monoclonal antibody for the detection of benzothiostrobin residue

Yulong Yuan; Xiude Hua; Ming Li; Wei Yin; Haiyan Shi; Minghua Wang

doi:10.1039/C4RA01845A

Development of a sensitive indirect competitive enzyme-linked immunosorbent assay based on the monoclonal antibody for the detection of benzothiostrobin residue

Yulong Yuan,^a Xiude Hua,^a Ming Li,^a Wei Yin,^a Haiyan Shi^a and Minghua Wang*^a

* Corresponding authors

^a Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University, Jiangsu Key Laboratory of Pesticide Science, Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing 210095, People's Republic of China
E-mail: wangmha@njau.edu.cn
Fax: +86 25 84395479
Tel: +86 25 84395479

Abstract

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on monoclonal antibodies (MAbs) for benzothiostrobin has been developed. The hapten of benzothiostrobin was synthesized and conjugated to bovine serum albumin and ovalbumin to generate an immunogen and a coating antigen. The immunogen was used to immunize BALB/c mice, resulting in anti-benzothiostrobin MAb. Under optimal conditions, the half maximal inhibition concentration (IC₅₀) and the limit of detection (IC₁₀) of the developed ic-ELISA were 7.55 and 0.428 μg L⁻¹, respectively. The cross-reactivity (CR) was less than 0.05% for the tested structural analogues and regarded as negligible except for pyraclostrobin, which exhibited a CR of 0.34%. The recoveries of benzothiostrobin ranged from 80.43 to 113.83% in environmental and agricultural samples, respectively, which conformed to the requirements for residue detection. The amount of benzothiostrobin detected by ic-ELISA in the samples was significantly (R² = 0.9894, y = 1.0867x + 0.0318) correlated with that detected by high-performance liquid chromatography (HPLC). The current study indicates that the established ic-ELISA is a potentially useful tool for detecting benzothiostrobin in environmental and agricultural samples.

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Publication details

The article was received on 03 Mar 2014, accepted on 15 May 2014 and first published on 16 May 2014

Article type: Paper

DOI: 10.1039/C4RA01845A

Citation: RSC Adv., 2014,4, 24406-24411

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Development of a sensitive indirect competitive enzyme-linked immunosorbent assay based on the monoclonal antibody for the detection of benzothiostrobin residue

Y. Yuan, X. Hua, M. Li, W. Yin, H. Shi and M. Wang, RSC Adv., 2014, 4, 24406
DOI: 10.1039/C4RA01845A
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Development of a sensitive indirect competitive enzyme-linked immunosorbent assay based on the monoclonal antibody for the detection of benzothiostrobin residue

Abstract

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Development of a sensitive indirect competitive enzyme-linked immunosorbent assay based on the monoclonal antibody for the detection of benzothiostrobin residue

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