Development and Optimization of an Indirect Enzyme‐Linked Immunosorbent Assay for Thiamphenicol

Hao, Kai; Guo, Shengda; Xu, Chuanlai

doi:10.1080/00032710600620443

Cited by 10 publications

(9 citation statements)

References 13 publications

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“…As shown in Table 1, several haptens had been designed to obtain the special antibodies against TAP, FF, and/or FFA in previous studies. Hao et al (2006) and Guo et al (2015) selected TAP as the starting material to generate a hapten and then to prepare antigens. Wu et al (2008), Luo et al (2009b), andFodey et al (2013) selected FFA as their haptens.…”

Section: Resultsmentioning

confidence: 99%

“…Synthesis of Hapten FF-SA The hapten FF succinate (FF-SA) was synthesized by coupling FF with succinic anhydride (SA) based on the modified method of Hao et al (2006). Briefly, a solution of FF (358 mg) in dichloromethane (15 mL) was added to succinic anhydride (159 mg) at room temperature.…”

Section: Synthesis Of Haptensmentioning

confidence: 99%

“…Recently, FF and TAP are even administered to fish by incorporation into feed for control of bacterial diseases (Hao et al 2006;Luo et al 2009a). Therefore, the need for an effective residue monitoring programme in place for FF and TAP is clear.…”

Section: Introductionmentioning

confidence: 99%

“…Hao et al (2006) developed an indirect ELISA to test TAP in eel using a polyclonal antibody (pAb) against TAP with IC 50 values 2.5 μg L −1 . Wu et al (Wu et al 2008) produced a pAb against FFA and developed an ic-ELISA to detect FFA with an IC 50 of 3.34 μg L −1 in muscle of swine and chicken.…”

Section: Introductionmentioning

confidence: 99%

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Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

Wang

Pan

et al. 2016

Food Anal. Methods

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To monitor the illegal use of florfenicol (FF) and thiamphenicol (TAP) in edible animal tissue and feed, a sensitive monoclonal antibody-based indirect competitive enzymelinked immunosorbent assay (ic-ELISA) has been developed with simple sample preparation and cleanup. The obtained monoclonal antibody (5F4) that has isotype IgG1 showed an IC 50 value of 0.21 μg L −1 for FF and 0.35 μg L −1 for TAP, respectively. It did not exhibit measurable cross-reactivity with other antibiotics. The limits of detection (LODs) for FF and TAP in a muscle matrix ranged from 0.07 to 0.14 μg kg −1 and in a feed matrix ranged from 2.9 to 5.2 μg kg −1 . The recoveries were 72.8 to 113.4 % with a coefficient of variation of less than 15 %. Good correlation between the ELISA and HPLC-MS/ MS results in the tissues tested demonstrated the reliability of our ic-ELISA. This ELISA is a useful tool for screening FF and TAP in edible animal tissue and feed.

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Section: Resultsmentioning

confidence: 99%

Section: Synthesis Of Haptensmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

Wang

Pan

et al. 2016

Food Anal. Methods

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“…The hapten FF-HS, which involved coupling FF with succinic anhydride, was synthesized in our laboratory following the method described by Hao, Guo, and Xu (2006). Specifically, FF (1.07 g, 3 mmol), succinic anhydride (0.6 g, 6 mmol) and pyridine (50 mL) were mixed and heated at 60°C for 4 h in a reflux condenser.…”

Section: Synthesis Of Hapten Ff-hsmentioning

confidence: 99%

Development of an ultrasensitive ic-ELISA and immunochromatographic strip assay for the simultaneous detection of florfenicol and thiamphenicol in eggs

Lei

Song

et al. 2017

Food and Agricultural Immunology

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An ultrasensitive monoclonal antibody-based gold nanoparticle immunochromatographic strip assay and indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) were developed to detect florfenicol (FF) and thiamphenicol (TAP) in egg samples. The ic-ELISA, with optimized pH, methanol content and sodium chloride content, exhibited an IC 50 value of 0.2 ng/mL for FF and 0.27 ng/mL for TAP, with the working range of 0.05-0.77 and 0.05-1.42 ng/mL, respectively. The optimized ic-ELISA showed negligible cross-reactivity with other phenols and broad-spectrum antibiotics. The recoveries in egg samples using the ic-ELISA ranged from 84% to 115% with a coefficient of variation of less than 5%. Based on this monoclonal antibody, a rapid and ultrasensitive immunochromatographic strip assay was developed with a cutoff value of 1 ng/mL for FF and TAP. Our results indicated that both developed methods were highly useful for screening FF and TAP in eggs.ARTICLE HISTORY

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Comparsion of an immunochromatographic strip with ELISA for simultaneous detection of thiamphenicol, florfenicol and chloramphenicol in food samples

Guo

Song

Liu

et al. 2015

Biomedical Chromatography

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Rapid and sensitive indirect competitive enzyme-linked immunosorbent assays (ic-ELISA) and gold nanoparticle immunochromatographic strip tests were developed to detect thiamphenicol (TAP), florfenicol (FF) and chloramphenicol (CAP) in milk and honey samples. The generic monoclonal antibody for TAP, FF and CAP was prepared based on a hapten [D-threo-1-(4-aminophenyl)-2- dichloroacetylamino-1,3-propanediol], and the haptenwas linked to a carrier protein using the diazotization method. After the optimization of several parameters (coating, pH, sodium chloride content and methanol content), the ic-ELISA was established. The quantitative working range for TAP was 0.11-1.36 ng/mL, with an IC50 of 0.39 ng/mL. The optimized ELISA showed cross-reactivity to CAP (300%) and FF (15.6%), with IC50 values of 0.13 and 2.5 ng/mL, respectively. The analytical recovery of TAP, FF and CAP in milk and honey samples in the ic-ELISA ranged from 81.2 to 112.9%. Based on this monoclonal antibody, a rapid and sensitive immunochromatographic test strip was also developed. This strip had a detection limit of 1 ng/mL for TAP, FF and CAP in milk and honey samples. Moreover, the test was completed within 10 min. Our results showed that the proposed ic-ELISA and immunochromatographic test strip method are highly useful screening tools for TAP, FF and CAP detection in milk and honey samples.

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Development and Optimization of an Indirect Enzyme‐Linked Immunosorbent Assay for Thiamphenicol

Cited by 10 publications

References 13 publications

Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

Development of an ultrasensitive ic-ELISA and immunochromatographic strip assay for the simultaneous detection of florfenicol and thiamphenicol in eggs

Comparsion of an immunochromatographic strip with ELISA for simultaneous detection of thiamphenicol, florfenicol and chloramphenicol in food samples

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