Determination of 3-Amino-2-oxazolidinone in Animal Tissue by an Enzyme-Linked Immunosorbent Assay and a Time-Resolved Fluoroimmunoassay

Abstract

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and a time-resolved fluoroimmunoassay (TR-FIA) for the determination of the furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) were developed. 3-Amino-2-oxazolidinone was detected using its derivative, 3-[[(2-nitrophenyl)-methylene]-amino]-2-oxazolidinone (NPAOZ). 3-([(3-Carboxyphenyl)methylene]-amino)-2-oxazolidinone (CPAOZ) was used as the immunizing hapten for the production of monoclonal antibodies against NPAOZ. The ic-ELISA had a 50 percent inhibition (IC 50 ) value of 0.82 microgram per liter and a detection limit of 0.11 microgram per liter. The recoveries from animal tissues spiked with AOZ from 0.2 to 5 micrograms per kilogram were 82.3-112.8 percent with coefficients of variation values of 3.5-12.5 percent. The time-resolved fluoroimmunoassay (TR-FIA) showed a 50 percent inhibition of 0.56 microgram per liter and a detection limit of 0.07 microgram per liter. The recoveries and coefficients of variation were 87.2-117.4 percent and 3.6-8.5 percent, respectively. The analysis of fortified samples by both methods provided similar results to those obtained by a standard high-performance liquid chromatographytandem mass spectrometry method. These results indicate the ic-ELISA and TR-FIA methods are suitable for the determination of 3-amino-2-oxazolidinone in animal tissue.

An immunochromatography test strip for rapid, quantitative and sensitive detection of furazolidone metabolite, 3-amino-2-oxazolidinone, in animal tissues

“…Gold chloride was provided by Sigma Company (St. Louis, MO, USA). Artificial antigen of CPAOZ-Bovine serum albumin (CPAOZ-BSA) conjugates, purifying anti-NPAOZ monoclonal antibodies (McAbs) was produced in our laboratory (Le & Yu, 2015). All other chemicals and organic solvents were of analytical grade or better.…”

“…The results showed that the ICTS is reliable for the quantitative detection of AOZ residue in animal tissue samples. Compared with the enzyme-linked immunosorbent assay method developed by Liu et al (2010), Zhu et al (2010), Jester et al (2014, and Le and Yu (2015), the ICTS could also help save time without losing accuracy and precision.…”

“…Many methods for the detection of AOZ have been developed, such as highperformance liquid chromatography with ultraviolet (Hu, Xu, & Yediler, 2007) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) (Hu et al, 2007;Valera-Tarifa, Plaza-Bolaños, Romero-González, Martínez-Vidal, & Garrido-Frenich, 2013;Wilasinee, Sutthivaiyakit, & Sutthivaiyakit, 2015;Zhang, Qiao, Chen, Wang, & Xia, 2016), high-performance liquid chromatography diode array detection (Fernando, Munasinghe, Gunasena, & Abeynayake, 2017) and enzyme-linked immunosorbent assays (Chang, Peng, Wu, Wang, & Yuan, 2008;Cheng et al, 2009;Cooper, Caddell, Elliott, & Kennedy, 2004;Franek et al, 2006;Jester, Abraham, Wang, EI Said, & Plakas, 2014;Le & Yu, 2015;Liu et al, 2010;Vass et al, 2005;Zhu et al, 2010). These analytical methods have many advantages of quantitative nature, precision and accuracy; however, they are time-consuming, expensive and unsuitable for analysis of large number of samples and also require expensive equipment and the procedure involves multiple steps.…”

“…Recently, several reports on colloidal gold-based ICTS methods have been published for the detection of AMOZ (Li et al, 2015), SEM (Tang, Xu, Wang, Xiang, & Yang, 2011;Yin, Liu, Song, Kuang, & Xu, 2015) and AHD (Tang, Xu, Liu, et al, 2011;Xu et al, 2009), but there have been no reports of colloidal gold-based ICTS method being used to measure AOZ in animal tissues. Based on the monoclonal antibody (McAb) against AOZ's derivates (NPAOZ) produced in our lab (Le & Yu, 2015), the aim of this study was to develop a rapid ICTS using colloidal gold-antibody probe for quantitative and sensitive detection of AOZ in animal tissues. Experimental results demonstrate that ICTS displayed the properties of high sensitivity, specificity, accuracy, precision and stability for AOZ detection.…”

Rapid and ultrasensitive detection of 3-amino-2-oxazolidinone in catfish muscle with indirect competitive enzyme-linked immunosorbent and immunochromatographic assays

“…Immunological analytical methods are also used to detect furazolidone residues, including enzyme-linked immunosorbent assays (ELISAs) and colloidal gold immunochromatographic assays. The immunoassay method has the advantages of rapidity, high sensitivity, high specificity, and high throughput (Gu, Liu, Song, Kuang, & Xu, 2016;Liu et al, 2010;Peng, Song, Liu, Kuang, & Xu, 2016), so many ELISAs have been developed to detect AOZ in aquatic animals, chicken, pork, beef, and eggs (Chang, Peng, Wu, Wang, & Yuan, 2008;Cheng et al, 2009;Cooper et al, 2004;Franek et al, 2006;Jester, Abraham, Wang, El Said, & Plakas, 2014;Le & Yu, 2015;Liu et al, 2010;Vass et al, 2005). Nonetheless, the ELISA is relatively cumbersome and requires expertise.…”