AUTHOR=Ren Xianfeng , Yue Xiaofeng , Mwakinyali Silivano Edson , Zhang Wen , Zhang Qi , Li Peiwu 

TITLE=Small Molecular Contaminant and Microorganism Can Be Simultaneously Detected Based on Nanobody-Phage: Using Carcinogen Aflatoxin and Its Main Fungal Aspergillus Section Flavi spp. in Stored Maize for Demonstration

JOURNAL=Frontiers in Microbiology

VOLUME=10

YEAR=2020

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.03023

DOI=10.3389/fmicb.2019.03023

ISSN=1664-302X

ABSTRACT=<p>Simultaneous detection technology has become a hot topic in analytical chemistry; however, very few reports on how to simultaneously detect small molecular contaminants and microorganisms have been in place. Aflatoxins are a group of highly toxic and carcinogenic compounds, which are produced mainly by <italic>Aspergillus flavus</italic> and <italic>Aspergillus parasiticus</italic> from section <italic>Flavi</italic> responsible for aflatoxin accumulation in stored cereals. Both aflatoxins and <italic>Aspergillus</italic> section <italic>Flavi</italic> were used to demonstrate the duplex real-time RCR method of simultaneously detecting small molecular contaminants and microorganisms. The detection of aflatoxins and <italic>Aspergillus</italic> section <italic>Flavi</italic> was carried out depending on the anti-idiotypic nanobody-phage V<sub>2–5</sub> and aflatoxin-synthesis related gene <italic>nor-1</italic> (=<italic>aflD</italic>), respectively. The quantitative standard curves for simultaneous detection of aflatoxins and <italic>Aspergillus</italic> section <italic>Flavi</italic> were constructed, with detection limits of 0.02 ng/ml and 8 × 10<sup>2</sup> spores/g, respectively. Naturally contaminated maize samples (<italic>n</italic> = 25) were analyzed for a further validation. The results were in good agreement between the new developed method and the referential methods (high-performance liquid chromatography and the conventional plating counts).</p>