Search

Herein, a novel visual method for detecting triazophos based on competitive bio-barcode immunoassay was described. The competitive immunoassay was established by gold nanoparticles (AuNPs), magnetic microparticle (MMPs) and triazophos, combined with biochip hybridization system to detect the residual of triazophos in water and apple. Because AuNPs carried many bio-barcodes, which hybridized with labeled DNA on the biochip, catalyzed signal amplification using silver staining was detected by grayscale values as well as the naked eye. Notably, the grayscale values decreases with increasing the concentrations of triazophos, and the color change weakened gradually. The detection range was in between 0.05 and 10 ng/mL and the minimum detection limit was set at 0.04 ng/mL. Percent recovery calculated from spiked water and apple samples ranged between 55.4 and 107.8% with relative standard deviations (RSDs) of 12.4-24.9%. It has therefore been shown that this protocol provides a new insight for rapid detection of small molecule pesticides in various matrices.

A fluorescence off-on method for the detection of Sorghum mosaic virus (SrMV) based on fluorescence resonance energy transfer (FRET) was developed in this study. The biosensor was realized by the adsorption of SrMV coat protein (CP) on gold nanoparticle (AuNP) and conjugation of anti-SrMV antibody (Ab) on cadmium telluride quantum dot (CdTe QD). The optimum quenching efficiency was about 50% obtained at a CdTe QD-Ab/AuNP-CP ratio of 1:7. Moreover, the feasibility of the developed biosensor was verified by the detection of purified CP and the limit of detection was estimated as 0.02 μg/mL. This strategy was also successfully applied to monitor SrMV CP in plant sap with a recovery rate between 97.7 and 107.4%. The developed biosensor is a simple, rapid, and efficient technique which does not need excessive washing and separation steps. [ABSTRACT FROM AUTHOR]

This study reports a novel bio-bar-code amplification competitive immunoassay for the detection of pesticide residue combined with a micro well plate. The sensing competitive immunoassay was made up of two types of particles: (1) the gold nanoparticles (AuNPs) with recognition elements (monoclonal antibody: McAb) and hundreds of thiolated single-strand oligonucleotide bar-codes; and (2) the magnetic microparticle (MMPs) with competition elements (hapten-OVA) for the target pesticide. Target pesticide and pesticide hapten competed with McAb labeled on the surface of AuNPs. The bar-code strands were released by dithiothreitol. Then a simple method based on micro well plate was used for the quantification of the bar-code strands. The bar-code strands with the half-complementary biotinylated DNA and half-complementary DNA-modified gold nanoparticles probe were immobilized onto the surface of a streptavidin-coated micro well plate, and the signal was amplified by silver enhancement. Triazophos (TAP), as a pesticide model, was detected by the proposed immunoassay. The linear range was from 2.5 × 10 −2 to 40.0 ng mL −1 , and LOD of the immunoassay was 1.96 × 10 −2 ng mL −1 . The correlation was further studied between this method and GC–MS analysis. Therefore, this method provided a promising approach for rapid detection and screening of pesticide residue. [ABSTRACT FROM AUTHOR]

A simple and highly sensitive immunoassay based on a competitive binding and bio-barcode amplification was designed for detection of small molecules, triazophos. The gold nanoparticles (AuNPs) were modified with monoclonal antibodies and 6-carboxyfluorescein labeled single-stranded thiol-oligonucleotides (6-FAM-SH-ssDNAs); the fluorescence of 6-FAM was quenched by AuNPs. Ovalbumin-linked haptens were coated on the bottom of microplate to compete with the triazophos in the sample for binding to the antibodies on the AuNP probes. The fluorescence intensity was inversely proportional to analyte concentration. Parameters of AuNP probes preparation and immune reaction were optimized. At the optimal conditions, the salting process was shortened to 1 h and 166 ± 9 ssDNAs were loaded onto a single AuNP. The competitive fluorescence bio-barcode immunoassay was performed on water, rice, cucumber, cabbage and apple samples. The linear range of the method was 0.01–20 μg L −1 and the limit of detection (LOD) was 6 ng L −1 . The recovery and relative standard deviations (RSDs) ranged from 85.0 to 110.3% and 9.4–17.4%, respectively. Good correlations were obtained between the results of the developed method and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In conclusion, it is suggested that the competitive fluorescent bio-barcode immunoassay had the potential to be used as a sensitive method for detection of a variety of small molecules in various samples. [ABSTRACT FROM AUTHOR]

The authors have developed a competitive immunoassay for the model pesticide triazophos (TRIAZ). The method is based on the use of only one monoclonal antibody immobilized on multi-labeled gold nanoparticle (AuNP) probes. The immunoassay uses two sets of probe. The first is a multi-labeled AuNP probe that carries monoclonal antibody (mcAb), single-stranded DNA (ssDNA) and horse radish peroxidase (HRP). The second is a magnetic microparticle (MMP) probe that is obtained by coating MMPs with ovalbumin coupled to TRIAZ. Free TRIAZ and MMP-immobilized TRIAZ compete for binding to the mcAb on the surface of the AuNPs. Because TRIAZ is a rather small molecule, it cannot be bound by two antibodies. The competitive immunoassay overcomes the limitations of small molecule detection using one kind of mcAb only. Sensitive transduction of the immunoreaction is accomplished by enzymatically catalyzed amplified. TRIAZ was quantified by a conventional ELISA and by the immunoassay presented here. Both method are highly sensitive and well reproducible. The new assay has a linear response in the 15 ng L to 40 μg L TRIAZ concentration range, and a 14 ng L limit of detection which make it more sensitive than the ELISA. The recovery rate in case of spiked samples ranges from 78.4 to 105%, and the RSD is <20%. A good correlation was further established between the results of the immunoassay and those of GC-MS analysis. [ABSTRACT FROM AUTHOR]