title: Rapid screening test for detection of oxytetracycline residues in milk using lateral flow assay.
abstract: A rapid, semi-quantitative lateral flow assay (LFA) was developed to screen the oxytetracycline (OTC) antibiotics residues in milk samples. In this study a competitive immuno-assay format was established. Colloidal gold nano-particles (GNP) were prepared and used as labelling material in LFA. Polyclonal antibodies were generated against OTC molecule (anti-OTC), purified and the quality was assessed by enzyme linked immuno sorbet assay. For the first time membrane components required for LFA in milk system was optimized. GNP and anti-OTC stable conjugate preparation method was standardized, and then these components were placed over the conjugate pad. OTC coupled with carrier protein was placed on test line; species specific secondary antibodies were placed on the control line of the membrane matrix. Assay was validated by spiking OTC to antibiotic free milk samples and results could be accomplished within 5min. without need of any equipment. The visual detection limit was 30ppb.
authors.affiliation: Dairy Chemistry Section, ICAR-National Dairy Research Institute, Southern Regional Station, Bengaluru, Karnataka 560030, India. Electronic address: laxmandcnaik@gmail.com.
authors.forename: Laxmana
authors.initials: L
authors.lastname: Naik
authors.affiliation: Dairy Chemistry Division, ICAR-National Dairy Research Institute, Karnal 132001, India.
authors.forename: Rajan
authors.initials: R
authors.lastname: Sharma
authors.affiliation: Dairy Chemistry Division, ICAR-National Dairy Research Institute, Karnal 132001, India.
authors.forename: Bimlesh
authors.initials: B
authors.lastname: Mann
authors.affiliation: Dairy Chemistry Division, ICAR-National Dairy Research Institute, Karnal 132001, India.
authors.forename: Kiran
authors.initials: K
authors.lastname: Lata
authors.affiliation: Animal Biochemistry Division, ICAR-National Dairy Research Institute, Karnal 132001, India.
authors.forename: Y S
authors.initials: YS
authors.lastname: Rajput
authors.affiliation: Dairy Chemistry Section, ICAR-National Dairy Research Institute, Southern Regional Station, Bengaluru, Karnataka 560030, India.
authors.forename: B
authors.initials: B
authors.lastname: Surendra Nath
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D017979:Gold Colloid; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D008892:Milk / Q000737:chemistry*; D010118:Oxytetracycline / Q000032:analysis*
chemical_list: D000900:Anti-Bacterial Agents; D017979:Gold Colloid; D010118:Oxytetracycline
keywords: Antibiotics; Gold nanoparticles; Lateral flow assay; Milk; Oxytetracycline