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

Abstract

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 Aspergillus flavus and Aspergillus parasiticus from section Flavi responsible for aflatoxin accumulation in stored cereals. Both aflatoxins and Aspergillus section Flavi were used to demonstrate the duplex real-time RCR method of simultaneously detecting small molecular contaminants and microorganisms. The detection of aflatoxins and Aspergillus section Flavi was carried out depending on the anti-idiotypic nanobody-phage V 2−5 and aflatoxin-synthesis related gene nor-1 (=aflD), respectively. The quantitative standard curves for simultaneous detection of aflatoxins and Aspergillus section Flavi were constructed, with detection limits of 0.02 ng/ml and 8 × 10 2 spores/g, respectively. Naturally contaminated maize samples (n = 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).

Fungi population metabolomics and molecular network study reveal novel biomarkers for early detection of aflatoxigenic Aspergillus species

“…In terms of time spent on analysis methods, our method takes 6-8 h from metabolites extraction to mass spectrometry analysis per 10 samples. Gene marker-based approach takes 4-5 h from DNA extraction to PCR test per 10 samples (Ren et al, 2020). The protein marker-based early detection method takes 18-24 h from protein extraction to immunoassay test per 10 samples (B.…”

Advances in phage display based nano immunosensors for cholera toxin

“…Ren et al. ( 170 ) devised an innovative detection platform that integrates PD-IPCR and RT-PCR, enabling simultaneous detection of multiple toxins in stored corn. PD-IPCR, an ultra-sensitive immunoassay for small-molecule biotoxins, was combined with real-time PCR assays capable of detecting various genes, including the cholera toxin gene.…”

Nanoanticuerpos: desarrollo biotecnológico y aplicaciones

“…Por otro lado, la efectividad que ejercen los Nac al aplicarlos se extiende hasta la industria agrícola y de los alimentos, donde se han desarrollado como herramientas de identificación y detección de las toxinas producidas por microorganismos como las bacterias y los hongos principalmente. Por ejemplo, las micotoxinas de Aspergillus flavus (Ren et al, 2020), y la producida por Bacillus thuringiensis, que se han utilizado como un bio-insecticida efectivo para el control de plagas agrícolas (Xu et al, 2017). Para el año 2022, en Estados Unidos se espera el lanzamiento del primer biofungicida con el nombre "Biofun-1", el cual combina el reconocimiento de los Nac y antifúngicos contra Botrytis cinerea (Njeru & Kusolwa, 2021).…”