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

Abstract

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.

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

“…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.…”

“…Fodey et al (2013 produced a panel of pAbs, which the most sensitive antibody could bind TAP (IC 50 = 0.75 μg L −1 ), FF (IC 50 = 0.61 μg L −1 ), and FFA (IC 50 = 0.93 μg L −1 ). Recently, Guo et al (2015) produced a monoclonal antibody (mAbs) that could bind TAP (IC 50 = 0.39 μg L −1 ), FF (IC 50 = 2.5 μg L −1 ), and CAP (IC 50 = 0.13 μg L −1 ) and developed ic-ELISA and gold nanoparticle immunochromatographic strip tests to detect TAP, FF, and CAP in milk and honey samples. Clearly, most reported ELISA methods for the detection of FF, TAP, and FFA residues are based on pAbs.…”

Development of a quantitative fluorescence-based lateral flow immunoassay for determination of chloramphenicol, thiamphenicol and florfenicol in milk

“…Considering that, it is necessary to establish a rapid, sensitive assay for the determination of CAP, TAP and FF residues in food samples. A number of analytical methods have been developed to detect CAP, TAP and FF; these include high-performance liquid chromatography (Xie et al, 2011), gas chromatographymass spectrometry (GC-MS) (Shen et al, 2009), liquid chromatography tandem mass spectrometry (LC-MS/MS) (Barreto, Ribeiro, Hoff, & Dalla Costa, 2016;Li, Hu, Huo, & Xu, 2006;Rezende, Filho, & Rocha, 2012), enzyme-linked immunosorbent assay (Fodey, Murilla, Cannavan, & Elliott, 2007;Liu et al, 2014; and lateral flow immunoassay (LFA) (Bai et al, 2013;Guo et al, 2015;Samsonova, Cannavan, & Elliott, 2012;Xu, Xu, Ma, Liu, et al, 2015). Compared with chromatographic methods, immunoassays are simple, rapid and do not require expensive equipment and professional operator.…”

Development of a multicomponent immunochromatographic test system for the detection of fluoroquinolone and amphenicol antibiotics in dairy products

“…ELISA protocols for the detection of CIP and CAP in food products have been described in various investigations . In comparison to ELISA, ICA is methodologically simple, does not require any equipment, and is rapid (the assay results can be visually assessed within 5–10 min) .…”