Generation and Characterization of an Anti-diclazuril Monoclonal Antibody and Development of a Diagnostic Enzyme-Linked Immunosorbent Assay for Poultry

Abstract

An anti-diclazuril monoclonal antibody (mAb) was developed for use in enzyme-linked immunosorbent assay (ELISA)-based detection of diclazuril with high sensitivity and specificity, which can be used to measure anti-coccidial drug residues. The anti-diclazuril mAb had a half-maximal inhibitory concentration of 0.449–0.517 ng/mL. The mAb cross-reactivity with toltrazuril, toltrazuril 18 sulfone, clozaril, monesin, madurmycin, and salinomycin was very minimal (< 0.1%). The detection limit of the ELISA using this mAb was 0.10 ng/mL and the sensitivity was 0.05 ng/mL. A standard curve generated in the range of 0.05–16.2 ng/mL had a linear correlation coefficient value of ≥ 0.99. The average recoveries of diclazuril from chicken and duck samples ranged from 85.0 to 102.5%.Intra- and inter-assay coefficients of variation ranged from 5.9 to 8.5% and 9.2 to 12.6%, respectively. Using the International Immunogenetics Information System®, the VH domain of the mAb was found to be encoded by an IGHV3 family gene and had the following complementarity determining region (CDR) sequences: GFTFSRY (CDR1), SRGGS (CDR2), and GDDNYAFAY (CDR3). The VL domain was encoded by an IGKV1 family gene and had the following CDR sequences: KSSQSLLNSRTRKNYLA (CDR1), WASTRES (CDR2), and KQSYNLHT (CDR3). This study provides a method to generate anti-diclazuril mAbs and determine their variable region sequences. The diagnostic ELISA developed using this mAb may drive additional studies on the monitoring and detection of food and veterinary drug residues.

Hapten designs based on aldicarb for the development of a colloidal gold immunochromatographic quantitative test strip

“…Mouse spleen cells were fused with myeloma cells, and the cell lines that could stably secrete anti-aldicarb monoclonal antibodies were screened through cell culturing, screening, and other steps ( 26 ). Ascites antibodies were prepared by in vivo induction, and the anti-aldicarb monoclonal antibody was purified using octanoic acid and saturated sulfuric acid following the ammonium method ( 27 ).…”

A Comparable icELISA and Lateral Flow Immunoassay for the Sensitive and Rapid Detection of 4,4′-Dinitrocarbanilide in Chicken

“…At present, the analytical methods for DNC in chicken, liver, and eggs mainly depend on instrumental analysis methods, including high-performance liquid chromatography, liquid chromatography–tandem mass spectrometry, and gas chromatography [ 7 ]. Although these methods are accurate, they tend to be complex in terms of sample preparation, and expensive and laborious in the detection procedure [ 8 ]. However, the analysis pattern of rapid detection is developing to be the alternative method for the rapid and immediate screening of DNC residues in samples [ 9 ].…”