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Lateral flow assays (LFA) are quick residue detection techniques used for screening of biological fluids such as milk, urine or blood and give quick and reliable results. In this study, a rapid and efficient lateral flow assay-based strip has been developed to detect sulfonamide residues in biological fluids. This kit has been designed on competitive format principle of LFA combined with nanotechnology. Primary antibodies were raised successfully in rabbits against hapten (Sulf-BSA) and purified by Octanoic acid-ammonium sulphate (OA-AS) sequential method. Total protein (IgG) content measured by nanodrop spectrophotometer was 8.53 mg/ml. Decreased absorbance values of anti-serum and purified serum preparations at same dilution during direct ELISA indicated that the purified serum antibodies certainly contained antibodies of interest (antisulfanilamide antibodies) in concentrated form. Characteristic peak of plasmon resonance of synthesized Gold nanoparticles (GNP; size: 65.19.8 nm and zeta potential:–21.6 mV) was observed at 520 nm with an absorbance of 0.40 followed by their conjugation with primary antibodies. On the nitrocellulose membrane the immunogen (Sulf-Ova) was smeared on the test line and secondary antibodies on the control line. GNP conjugates were impregnated on the conjugate pad. Strips were then run with 50, 100,150 and 200 ppb of sulphanilamide concentrations. At 50 ppb concentration, the sulphanilamide could not be detected whereas at 100, 150 and 200 ppb the sulphanilamide was detected in the samples (Milk and Buffer). The developed assay could detect maximum residue limit values of sulfonamides in milk. The visual detection was achieved by using GNPs. Duration of analysis was 8 to 15 min. These strips are easy to use and can be successfully used for screening of sulfonamides in biological fluids.
Faqir Muhammad, Nida Noreen, Bilal Aslam. (2021) DEVELOPMENT OF USER-FRIENDLY DIAGNOSTIC KIT FOR DETECTION OF SULFONAMIDE RESIDUES IN BIOLOGICAL FLUIDS, Pakistan Journal of Agricultural Sciences, Volume 58, Issue 2.
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