Anti-idiotypic nanobody-phage based real-time immuno-PCR for detection of hepatocarcinogen aflatoxin in grains and feedstuffs

Anal Chem. 2014 Nov 4;86(21):10841-6. doi: 10.1021/ac5029424. Epub 2014 Oct 14.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aflatoxins / analysis*
  • Aflatoxins / toxicity
  • Animal Feed / analysis*
  • Antibodies, Anti-Idiotypic / immunology*
  • Base Sequence
  • Carcinogens / analysis
  • Carcinogens / toxicity
  • DNA Primers
  • Edible Grain / chemistry*
  • Liver Neoplasms / chemically induced
  • Real-Time Polymerase Chain Reaction / methods*
  • Single-Domain Antibodies / immunology*

Substances

  • Aflatoxins
  • Antibodies, Anti-Idiotypic
  • Carcinogens
  • DNA Primers
  • Single-Domain Antibodies