Preparation of anti-pefloxacin antibody and development of an indirect competitive enzyme-linked immunosorbent assay for detection of pefloxacin residue in chicken liver.
Pefloxacin has been increasingly used in veterinary medicine to treat microbial infections. To avoid using a labor-intensive instrumental method to detect the residue of pefloxacin in food, a simple and convenient indirect competitive enzyme-linked immunosorbent assay method has been developed in this study. The antibody generated from immunogen cationized bovine serum albumin-pefloxacin showed high sensitivity toward pefloxacin with an IC50 value of 6.7 ppb in buffer and was suitable for a screening assay to detect the residue of pefloxacin in food products. The antibody has been assessed using rapid enzyme immunoassays to exploit its specificity. The antibody prepared shows cross-reactivity with a few other (fluoro)quinolones including fleroxacin (116%), enrofloxacin (88%), and ofloxacin (10%). The assay measured drug residue in chicken liver spiked with pefloxacin with an interassay coefficient of variation of 13.6% or less and an intra-assay coefficient of variation of 10.9% or less. The average recovery rates at 0.5, 5, 10, 50, and 100 ppb were in the range of 86-106% for interassay and in the range of 87-103% for intra-assay, respectively.
Lu
Shengxin
S
The School of Chemistry and Chemical Engineering, Shandong University, Jinan, Shandong 250100, People's Republic of China.
Zhang
Yulan
Y
Liu
Jingting
J
Zhao
Chengbiao
C
Liu
Wei
W
Xi
Rimo
R
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001667:Binding, Competitive; D002645:Chickens; D004350:Drug Residues / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008099:Liver / Q000737:chemistry*; D008297:Male; D015366:Pefloxacin / Q000032:analysis* / Q000276:immunology*; D011817:Rabbits; D012680:Sensitivity and Specificity
D000906:Antibodies; D015366:Pefloxacin