title: Anti-idiotypic nanobody-phage based real-time immuno-PCR for detection of hepatocarcinogen aflatoxin in grains and feedstuffs.
abstract: Aflatoxins are a group of extremely toxic small molecules that have been involved in human hepatic and extrahepatic carcinogenesis as causative agents. Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-products base on a M13 phage containing aflatoxin anti-idiotypic nanobody and its encoding DNA which was used to design the specific primers. The limit of detection (LOD) of the assay is 0.02 ng/mL, which exhibits a 4-fold improvement over traditional phage ELISA. The developed method was successfully validated with the samples of corn, rice, peanut, and feedstuff, which are major aflatoxin-contaminated agri-products. And the recoveries were from 77.05 to 122.16%. For further validation, the developed assay was also compared with a reference HPLC method for the analysis of aflatoxins in corn and peanuts, and concordant results (R(2) = 0.991) were obtained. In this context, this study provides a novel opportunity to analyze aflatoxins in agri-products.
authors.affiliation: Oil Crops Research Institute, Chinese Academy of Agricultural Sciences , Wuhan, 430062, People's Republic of China.
authors.forename: Jiawen
authors.initials: J
authors.lastname: Lei
authors.forename: Peiwu
authors.initials: P
authors.lastname: Li
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.forename: Yanru
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Zhang
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Ding
authors.forename: Wen
authors.initials: W
authors.lastname: Zang
mesh_terms: D000348:Aflatoxins / Q000032:analysis* / Q000633:toxicity; D000821:Animal Feed / Q000032:analysis*; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D001483:Base Sequence; D002273:Carcinogens / Q000032:analysis / Q000633:toxicity; D017931:DNA Primers; D002523:Edible Grain / Q000737:chemistry*; D008113:Liver Neoplasms / Q000139:chemically induced; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D061905:Single-Domain Antibodies / Q000276:immunology*
chemical_list: D000348:Aflatoxins; D000888:Antibodies, Anti-Idiotypic; D002273:Carcinogens; D017931:DNA Primers; D061905:Single-Domain Antibodies