Design and synthesis of immunoconjugates and development of an indirect ELISA for rapid detection of 3, 5-dinitrosalicyclic Acid hydrazide.
In this study novel immunoconjugates were designed, synthesized and then used to develop a rapid, specific and sensitive indirect ELISA method to directly detect residues of 3,5-dinitrosalicyclic acid hydrazide (DNSH), a toxic metabolite of nifursol present in chicken tissues. The hapten DNSHA was first designed and used to covalently couple to BSA to form an immunogen which was immunized to rabbits to produce a polyclonal antibody against DNSH. Furthermore, a novel 3,5-dinitrosalicylic acidovalbumin (DNSA-OVA) immunoconjugate structurally different from DNSHA-OVA was designed and used as a "substructural coating antigen" to improve the sensitivity of an indirect ELISA analysis for a direct DNSH detection. Based on the "substructural coating antigen" concept, an optimized indirect ELISA method was established that exhibited good specificity and high sensitivity for detecting DNSH, with a cross-reactivity of less than 0.1% (excluding the parent compound nifursol), IC(50) of 0.217 nmol/mL and detection limit of 0.018 nmol/mL. Finally, a simple and efficient analysis of DNSH samples in chicken tissues showed that the average recovery rate of the indirect ELISA analysis was 82.3%, with the average coefficient of variation 15.9%. Thus, the developed indirect ELISA method exhibited the potential for a rapid detection of DNSH residues in tissue.
Shen
Yu-Dong
YD
College of Food Science, South China Agricultural University, Guangzhou 510642, P. R. China.
Zhang
Shi-Wei
SW
Lei
Hong-Tao
HT
Wang
Hong
H
Xiao
Zhi-Li
ZL
Jiang
Yue-Ming
YM
Sun
Yuan-Ming
YM
D000818:Animals; D000918:Antibody Specificity; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006241:Haptens / Q000737:chemistry; D006834:Hydrazines / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology; D008460:Meat / Q000032:analysis*; D015394:Molecular Structure; D009581:Nitrofurans / Q000737:chemistry; D015203:Reproducibility of Results
D006241:Haptens; D006834:Hydrazines; D018796:Immunoconjugates; D009581:Nitrofurans; C029424:hydrazine; C032119:nifursol