Ultrasensitive fluorometric oligonucleotide immunoassay for the simultaneous and efficient detection of two mycotoxins in agricultural products.

An ultrasensitive fluorometric oligonucleotide immunoassay (UFOIA) based on a fluorometric oligonucleotide and magnetic separation was proposed for the simultaneous detection of two mycotoxins. Two kinds of magnetic nanoparticle (MNP) probes and their corresponding fluorometric oligonucleotide probes were prepared. After the immune reaction, Cy5-linked and 6-FAM-linked oligonucleotides were dissociated and applied to detect fluorescence signals simultaneously. Under optimal conditions, the detection ranges of the UFOIA were in the range of 0.654-1438.8 pg mL-1 for zearalenone (ZEN) and 0.215-3190.1 pg mL-1 for aflatoxin B1 (AFB1). The limits of detection (LODs) were 0.378 pg mL-1 for ZEN and 0.043 pg mL-1 for AFB1, which showed improved sensitivities of 529-fold and 112-fold compared to those from the ELISA. The positive results of the UFOIA for authentic agricultural products were highly correlated with those from LC-MS/MS. The specificity, accuracy, precision and reliability of the UFOIA are well demonstrated. The proposed UFOIA method achieved the simultaneous and ultrasensitive detection of mycotoxins at the pg mL-1 level, which was a considerable improvement. This study might provide an alternative approach for detecting multi-component contamination equipped with the notable highlights of ultrasensitivity, simultaneity, simplicity, high efficiency and a low background signal.

Ultrasensitive monitoring strategy of PCR-like levels for zearalenone contamination based DNA barcode.

BACKGROUND: The ultrasensitive monitoring strategy of zearalenone (ZEN) is essential and desirable for food safety and human health. In the present study, a coupling of gold nanoparticles-DNA barcode and direct competitive immunoassay-based real-time polymerase chain reaction signal amplification (RT-IPCR) for ZEN close to the sensitivity of PCR-like levels is described and evaluated. RESULTS: The RT-IPCR benefited from the use of a DNA barcode and RT-PCR detection strategy, thus resulting in ultrasensitive and simple detection for ZEN. Under the optimal RT-IPCR, the linear range of detection was from 0.5 to 1000 pg mL-1 and the limit of detection was 0.5 pg mL-1 , which was 400-fold lower than the enzyme-linked immunosorbent assay. The detection procedure was simplified and the detection time was shortened. The specificity, accuracy and precision of the RT-IPCR confirmed a high performance. ZEN-positive contamination levels were from 0.056 to 152.12 ng g-1 by the RT-IPCR, which was demonstrated to be highly reliable by liquid chromatography-tandem mass spectrometry. CONCLUSION: The proposed RT-IPCR could be used as an alternative for detecting ZEN with satisfactory ultrasensitivity, simplicity, low cost and high-throughput. The present study could provide a strategy for the ultrasensitive detection of the small molecule with a simple and practical approach, which has significant appeal and application prospects.

Contamination of Zearalenone from China in 2019 by a Visual and Digitized Immunochromatographic Assay.

Zearalenone (ZEN) is a prevalent mycotoxin that needs intensive monitoring. A semi-quantitative and quantitative immunochromatographic assay (ICA) was assembled for investigating ZEN contamination in 187 samples of cereal and their products from China in 2019. The semi-quantitative detection model had a limit of detection (LOD) of 0.50 ng/mL with visual judgment and could be completely inhibited within 5 min at 3.0 ng/mL ZEN. The quantitative detection model had a lower LOD of 0.25 ng/mL, and ZEN could be accurately and digitally detected from 0.25-4.0 ng/mL. The ICA method had a high sensitivity, specificity, and accuracy for on-site ZEN detection. For investigation of the authentic samples, the ZEN-positive rate was 62.6%, and the ZEN-positive levels ranged from 2.7 to 867.0 ng/g, with an average ZEN-positive level being 85.0 ng/g. Of the ZEN-positive samples, 6.0% exceeded the values of the limit levels. The ZEN-positive samples were confirmed to be highly correlated using LC-MS/MS (R2 = 0.9794). This study could provide an efficiency and accuracy approach for ZEN in order to achieve visual and digitized on-site investigation. This significant information about the ZEN contamination levels might contribute to monitoring mycotoxin occurrence and for ensuring food safety.

Zearalenone Contamination in Corn, Corn Products, and Swine Feed in China in 2016-2018 as Assessed by Magnetic Bead Immunoassay.

In total, 405 samples of corn, corn products, and swine feed from China in 2016-2018 were surveyed for zearalenone (ZEN) contamination using a magnetic bead immunoassay-coupled biotin-streptavidin system (BAS-MBI). The developed BAS-MBI had a limit of detection (LOD) of 0.098 ng mL-1, with half-maximal inhibition concentration (IC50) of 0.71 ng mL-1 in working buffer, and an LOD of 0.98 ng g-1; the detection range was from 0.98 to 51.6 ng g-1 in authentic agricultural samples. The BAS-MBI has been demonstrated to be a powerful method for the rapid, sensitive, specific, and accurate determination of ZEN. The ZEN positivity rate reached the highest level of 40.6% in 133 samples in 2016; ZEN levels ranged from 1.8 to 1100.0 ng g-1, with an average level of 217.9 ng g-1. In 2017, the ZEN positivity rate was the lowest at 24.5% in 143 samples; ZEN levels ranged from 1.1 to 722.6 ng g-1, with an average of 166.7 ng g-1. In 2018, the ZEN positivity rate was 31.8% in 129 samples; ZEN levels ranged from 1.3 to 947.8 ng g-1, with an average of 157.0 ng g-1. About 20% of ZEN-positive samples exceeded maximum limit levels. An alternative method of ZEN detection and a valuable reference for ZEN contamination in corn and its related products in China are provided. This survey suggests the need for prevention of serious ZEN contamination, along with management for food safety and human health.

Nucleophilic addition-triggered lanthanide luminescence allows detection of amines by Eu(thenoyltrifluoroacetone)3.

Herein, a novel fluorescent indicator for the real-time monitoring of amines is described. This probe contains a complex of europium-(thenoyltrifluoroacetone)(3) (Eu(TTA)(3)) that efficiently reacts with primary and secondary amines. The electron-withdrawing trifluoroacetyl undergoes a nucleophilic addition with amines, and the complex was used to selectively detect BuNH(2) and Et(2) NH (quenching concentration for BuNH(2): 10(-4) M, for Et(2)NH: 1.2 × 10(-3) M) by monitoring emission; no changes were observed in the emission spectrum of Eu(TTA)(3) in the presence of Et(3)N, [Bu(4)N]Cl, or PhNH(2) in aqueous solution (THF/H(2)O = 1:1). The ratio of emission intensity to amine concentration was linear by the least-squares fitting method.