A competitive sensitive bio-barcode immunoassay based
on bimetallic
nanozyme (Au@Pt: gold@platinum) catalysis has been designed for the
detection of the pesticide parathion. Gold nanoparticles (AuNPs) were
modified with single-stranded thiol oligonucleotides (ssDNAs) and
monoclonal antibodies (mAbs) to form AuNP probes; magnetic nanoparticles
(MNPs) were coated with ovalbumin (OVA)–parathion haptens as
MNP probes, and bimetallic nanozyme (Au@Pt) nanoparticles functionalized
with the complementary thiolated ssDNA were used as Au@Pt probes.
The Au@Pt probes reacted with the AuNP probes through complementary
base pairing. Further, parathion competed with MNP probes to bind
the mAbs on the AuNP probes. Finally, the complex system was separated
by a magnetic field. The released Au@Pt probes catalyzed a chromogenic
system consisting of teramethylbenzidine (TMB). The bimetallic nanozyme-based
bio-barcode immunoassay was performed on rice, pear, apple, and cabbage
samples to verify the feasibility of the method. The immunoassay exhibited
a linear response from 0.01 to 40 μg·kg–1, and the limit of detection (LOD) was 2.13 × 10–3 μg·kg–1. The recoveries and relative
standard deviations (RSDs) ranged from 73.12 to 116.29% and 5.59 to
10.87%, respectively. The method was found to correlate well with
data obtained by liquid chromatography–tandem mass spectrometry
(LC–MS/MS). In conclusion, this method exhibits potential as
a sensitive alternative method for the detection of a variety of pesticides,
ensuring the safety of fruits and vegetables in agriculture.
An ultrasensitive bio-barcode competitive
immunoassay method based
on droplet digital polymerase chain reaction (ddPCR) was developed
for the determination of triazophos. Gold nanoparticles (AuNPs) were
coated with monoclonal antibodies (mAbs) and complementary double-stranded
DNA (dsDNA), which included bio-barcode DNA and thiol-capped DNA.
Magnetic nanoparticle (MNP) probes were constructed by modifying the
MNPs with ovalbumin–hapten conjugates (OVA–hapten).
The target pesticide and OVA–hapten on the surface of the MNP
probes competed with the AuNP probes simultaneously, and then the
bio-barcode DNA was released for quantification by ddPCR. The concentration
of released DNA was inversely proportional to the concentration of
pesticide to be tested. Under the optimum conditions, the competitive
immunoassay exhibited a wide linear range of 0.01–20 ng/mL
and a low detection limit of 0.002 ng/mL. Spike recovery tests were
carried out using apple, rice, cabbage, and cucumber samples to verify
the feasibility of the method. The recovery and relative standard
deviations (RSDs) of the technique ranged from 76.9 to 94.4% and from
10.8 to 19.9%, respectively. To further validate the results, a linear
correlation analysis was performed between the proposed method and
liquid chromatography–tandem mass spectrometry (LC–MS/MS).
Consequently, the bio-barcode immunoassay based on nanoparticles and
ddPCR, an ultrasensitive method, showed great potential for the determination
of target pesticides in real samples.
A rapid, ultra-sensitive and simple fluorescence immunoassay (FLISA) on CdSe/ZnS Quantum dots (QDs) for recognition pesticide triazophos have been established. In this assay, the immunoassay uses (CdSe/ZnS) Quantum dots (QDs) as a probe that carries monoclonal antibody (mAb). The free triazophos and OVA-haptens were competed for binding the mAb on the surface of the QDs. Therefore, the concentration of TRIAZ can get by calculation of the detection of fluorescence value. The assay has a linear response in the 10 ng L −1-25 μg L −1 TRIAZ concentration range, and limit of detection (IC 10) was 0.508 ng L −1. The recovery rate in the case of spiked samples ranges from 82.6% to 96.6%, and the RSD is <20%. FLISA can accurately and sensitively screen out triazophos in representative actual samples that contain trace levels of pesticides, it can play a prominent part in agricultural product and environmental detection.
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