A Dynamic and Pseudo-Homogeneous MBs-icELISA for the Early Detection of Aflatoxin B1 in Food and Feed.

Aflatoxin B1 (AFB1) is one of the most toxic and harmful fungal toxins to humans and animals, and the fundamental way to prevent its entry into humans is to detect its presence in advance. In this paper, the monoclonal antibody mAbA2-2 was obtained via three-step sample amplification and multi-concentration standard detection using a subcloning method based on the limited dilution method with AFB1 as the target. A dynamic and pseucdo-homogeneous magnetic beads enzyme-linked immunosorbent assay (MBs-icELISA) was established using the prepared antibody as the recognition element and immunomagnetic beads as the antigen carrier. The MBs-icELISA showed good linear correlation in the concentration range of 0.004-10 ng/mL with R2 = 0.99396. The limit of detection (LOD) of the MBs-icELISA for AFB1 was 0.0013 ng/mL. This new ELISA strategy significantly shortened AFB1 detection time through improved sensitivity compared to the conventional ELISA method.

Novel Broad-Spectrum Aptamer Used to Build Fe-Co Nanoporous Carbon Highly Sensitive Sensing Platform for the Detection of Multiple Organophosphorus Pesticides.

In this work, broad-spectrum aptamers for organophosphorus pesticides (OPs) were obtained by alternate target systematic evolution of ligands by exponential enrichment screening. The secondary and tertiary structure analyses of the aptamer inferred that the neck-loop structure formed a G-triplex structure with the target. In addition, optimization of the sheared aptamer resulted in a stronger affinity (Kd = 86.74 nM), which was increased by 2 orders of magnitude compared to similar aptamers. A novel electrochemical biosensor was prepared by modifying an aptamer labeled with an electroactive substance (methylene blue) on the surface of nanoporous carbon containing Fe-Co (Fe-Co/NPC). When a target bound to the aptamer, a G-triplex structure was formed close to the electrode surface. The aptamer phosphate backbone labeled with methylene blue enhanced the electron-transfer efficiency and resulted in signal changes. The biosensor exhibited an excellent sensitivity (7.32 fM) and a wide detection range (1 × 10-13 to 1 × 10-3 M) for OPs under optimal conditions, enabling simultaneous detection of multiple OPs in vegetables.

Multiplex strategy electrochemical platform based on self-assembly dual-site DNA tetrahedral scaffold for one-step detection of diazinon and profenofos.

In the work, based on self-assembly dual-site DNA tetrahedral scaffold (DTS), thionine (Thi), and 6-(Ferrocenyl)hexanethiol (Fc6S), a multiplex strategy electrochemical platform was fabricated for the simultaneous detection of profenofos (PFF) and diazinon (DZN). Thi and Fc6S were used to label aptamers for the synthesis of probes respectively. Notably, Thi and Fc6S engendered recognizable DPV peaks at different potentials to achieve simultaneous detection of PFF and DZN. In addition to increasing the conductivity of the electrode, the combination of carboxylic acid functionalized multi-walled carbon nanotubes and ferroferric oxide nanoparticles could also increase its higher specific surface area of the electrode interface to adsorb more DTS. Because of the mechanical rigidity of the DTS, the DTS could keep a complementary chain upright and provide more binding sites for aptamers, the binding efficiency between the complementary chain and 2 binding aptamers could be improved. Comparing the aptasensors performance of single-strand DNA with that of the DTS with complementary strands, the benefits of the DTS were highlighted in this system. Under optimal conditions, the detection limits of PFF and DZN were both 3.33 pg/mL and the detection ranges were both 1.00 × 101-1.00 × 107 pg/mL. Meanwhile, the recoveries of PFF and DZN were 87.15%-117.34% and 91.20%-114.19%, respectively. The aptasensor could realize the simultaneous detection of PFF and DZN in vegetables. Furthermore, the aptasensor also had good stability and selectivity. This strategy could provide a good reference for developing effective aptasensors for the simultaneous detection of other small molecules and toxins.

Interference-resistant aptasensor with tetrahedral DNA nanostructure for profenofos detection based on the composites of graphene oxide and polyaniline.

In this work, an interference-resistant electrochemical aptasensor that could detect profenofos in vegetables was constructed based on complexes of graphene oxide and polyaniline (GO@PANI) and gold nanoparticles-tetrahedral DNA nanostructure (Au-TDN). Compared with a single chain aptamer, the tetrahedral DNA nanostructure is highly stable and allows the aptamer on this structure to stand in a highly ordered position on an electrode surface. Moreover, the AuNPs are biocompatible and can protect the activity of the aptamer, which can improve the assembly success rate of Au-TDN. Besides, the conductivity of PANI had been tremendously enhanced thanks to the existence of GO, which improved the dispersion of PANI. The GO@PANI was prepared by a chemical synthesis method, which had a large surface area and was able to adsorb many Au-TDN. Under optimal working parameters, the constructed aptasensor exhibited good electrochemical sensing performance with a detection limit of 10.50 pg/mL and a linear range of 1.0 × 102-1.0 × 107 pg/mL. In addition, it was employed in detecting profenofos in vegetables with a good recovery rate of 90.41-116.37 %. More importantly, the aptasensor also has excellent stability and high selectivity. This study provides a promising method to avoid interference in the detection of profenofos by sensors.

Novel colorimetric aptasensor based on MOF-derived materials and its applications for organophosphorus pesticides determination.

For the visual detection of four organophosphorus pesticides (OPs), a colorimetric aptasensor was developed based on aptamer-mediated bimetallic metal-organic frameworks (MOFs) nano-polymers. Fe-Co magnetic nanoparticles (MNPs) and Fe-N-C nanozymes were prepared based on pyrolytic reaction, and were labeled with broad spectrum aptamers and complementary chains of organophosphorus pesticides respectively. The hybridization of aptamers and complementary chains led to the formation of nano-polymers. In the presence of target pesticides, they competed with complementary chains for aptamers on Fe-Co MNPs, resulting in a large number of Fe-N-C nanozymes signal labels being released into the supernatant. Fe-N-C nanozymes showed similar activity to peroxidase and catalyzed the 3,3',5,5'-tetramethylbenzidine-hydrogen peroxide (TMB-H2O2) color system to turn the solution blue-green under mild conditions. The magnetic probes had good selectivity and sensitivity, and were easily separated by magnetic absorption. The sensor functioned well under optimal conditions, demonstrating good stability and specificity for four pesticides: phorate, profenofos, isocarbophos and omethoate, and the detection limits of four pesticides were as low as 0.16 ng/mL, 0.16 ng/mL, 0.03 ng/mL and 1.6 ng/mL respectively, and the recovery rate of OPs residue in vegetable samples was satisfactory. The work described here provided a simple, rapid and sensitive way to construct a biosensor.

Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for visual/sensitive detection of kanamycin.

A dual-model colorimetric and electrochemical aptasensor was designed using a large number of G-quadruplexes generated by rolling circle amplification (RCA). Specific binding between target and aptamer during RCA yielded large numbers of G-quadruplexes. A colorimetric sensor was fabricated based on the interaction between the G-quadruplex and hemin, which altered the 3,3',5,5'-Tetramethylbenzidine (TMB)-catalyzed color reaction and facilitated the visual and semi-quantitative detection of kanamycin. An electrochemical sensor was constructed based on the strong interaction between the G-quadruplex and the methylene blue electrical signal molecule. Combining nanocomposites multi-walled carbon nanotubes-chitosan/gold nanoparticles (MWCNTs-CS/AuNPs) and RCA realized double-amplified electrochemical signals. Under optimized conditions, a linear relationship was obtained as the logarithm of different concentrations of kanamycin (KAN). The colorimetric aptasensor had a linear range of 1 × 102 nM to 1 × 103 nM with a detection limit of 1.949 nM. The electrochemical aptasensor had wider a linear range from 1 × 10-3 nM to 2.5 × 103 nM and a lower detection limit of 0.333 pM. The sensor combined the advantages of simple colorimetric visualization with the ultra-precision of electrochemical methods. Aptasensor showed good specificity and prevented interference. Furthermore, the prepared dual-model aptasensor facilitated the practical monitoring of KAN in milk.

Broad-specificity time-resolved fluorescent immunochromatographic strip for simultaneous detection of various organophosphorus pesticides based on indirect probe strategy.

The massive use of organophosphorus pesticides (OPs) poses a great threat to food safety, human health and environmental protection. As there are many kinds of pesticides, their detection is facing a severe challenge. The simultaneous detection of multiple organophosphorus pesticides in one test is a problem to be solved at present. In this paper, a time-resolved fluorescent immunochromatographic (TRFIA) strip is prepared by using broad-specificity antibodies (Abs) of OPs as the recognition element. Abs were connected to europium oxide latex microspheres using sheep anti-mouse antibodies (SaMIgG) to form an indirect probe. This strategy could effectively realize signal amplification, and could save the amount and protect the activity of Abs. After the detection, the color change of the test line (T-line) was observed to make qualitative judgment under UV-light (365 nm). Then, the images of the positive sample were analyzed by using ImageJ to complete the quantitative detection. Under optimal construction and operating conditions, the limit of detection of the strip could reach 0.53 ng g-1. And the TRFIA strip performed well in the additive test of vegetable samples. It is inexpensive to prepare, convenient to carry, and easy to operate. More importantly, it improves the detection efficiency and meets the needs of rapid field testing of a large number of samples.