scholarly journals A Novel Graphene Oxide-Based Aptasensor for Amplified Fluorescent Detection of Aflatoxin M1 in Milk Powder

Sensors ◽  
2019 ◽  
Vol 19 (18) ◽  
pp. 3840 ◽  
Author(s):  
Xiaodong Guo ◽  
Fang Wen ◽  
Qinqin Qiao ◽  
Nan Zheng ◽  
Matthew Saive ◽  
...  

In this paper, a rapid and sensitive fluorescent aptasensor for the detection of aflatoxin M1 (AFM1) in milk powder was developed. Graphene oxide (GO) was employed to quench the fluorescence of a carboxyfluorescein-labelled aptamer and protect the aptamer from nuclease cleavage. Upon the addition of AFM1, the formation of an AFM1/aptamer complex resulted in the aptamer detaching from the surface of GO, followed by the aptamer cleavage by DNase I and the release of the target AFM1 for a new cycle, which led to great signal amplification and high sensitivity. Under optimized conditions, the GO-based detection of the aptasensor exhibited a linear response to AFM1 levels in a dynamic range from 0.2 to 10 μg/kg, with a limit of detection (LOD) of 0.05 μg/kg. Moreover, the developed aptasensor showed a high specificity towards AFM1 without interference from other mycotoxins. In addition, the technique was successfully applied for the detection of AFM1 in infant milk powder samples. The aptasensor proposed here offers a promising technology for food safety monitoring and can be extended to various targets.

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1207
Author(s):  
Hong Jae Cheon ◽  
Quynh Huong Nguyen ◽  
Moon Il Kim

Inspired by the active site structure of natural horseradish peroxidase having iron as a pivotal element with coordinated histidine residues, we have developed histidine coated magnetic nanoparticles (His@MNPs) with relatively uniform and small sizes (less than 10 nm) through one-pot heat treatment. In comparison to pristine MNPs and other amino acid coated MNPs, His@MNPs exhibited a considerably enhanced peroxidase-imitating activity, approaching 10-fold higher in catalytic reactions. With the high activity, His@MNPs then were exploited to detect the important neurotransmitter acetylcholine. By coupling choline oxidase and acetylcholine esterase with His@MNPs as peroxidase mimics, target choline and acetylcholine were successfully detected via fluorescent mode with high specificity and sensitivity with the limits of detection down to 200 and 100 nM, respectively. The diagnostic capability of the method is demonstrated by analyzing acetylcholine in human blood serum. This study thus demonstrates the potential of utilizing His@MNPs as peroxidase-mimicking nanozymes for detecting important biological and clinical targets with high sensitivity and reliability.


2021 ◽  
Vol 11 (11) ◽  
pp. 1774-1780
Author(s):  
Shanji Fan ◽  
Hong Huang ◽  
Hong Chen ◽  
Jiachi Xu ◽  
Zecheng Hu ◽  
...  

A CdS nanocrystal enhanced TiO2 nanotubes (CdS@TiO2 NATs) photoelectrode was prepared via successive ionic layer adsorption and reaction (SILAR) of CdS on the surface of TiO2 NATs. A HS-aptamer owing a specific binding toward cytochrome c was modified onto the CdS@TiO2 NATs, which resulting a decrease in the photoelectrical current intensity. Cytochrome c is therefore quantified based on the decrease in photoelectrical current. High specificity and high sensitivity were obtained with a linear range from 3 pM to 80 nM, and a limit of detection of 2.53 pM.


2021 ◽  
Vol 8 ◽  
Author(s):  
Alfredo Garcia-Venzor ◽  
Bertha Rueda-Zarazua ◽  
Eduardo Marquez-Garcia ◽  
Vilma Maldonado ◽  
Angelica Moncada-Morales ◽  
...  

As to date, more than 49 million confirmed cases of Coronavirus Disease 19 (COVID-19) have been reported worldwide. Current diagnostic protocols use qRT-PCR for viral RNA detection, which is expensive and requires sophisticated equipment, trained personnel and previous RNA extraction. For this reason, we need a faster, direct and more versatile detection method for better epidemiological management of the COVID-19 outbreak. In this work, we propose a direct method without RNA extraction, based on the Loop-mediated isothermal amplification (LAMP) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated protein (CRISPR-Cas12) technique that allows the fast detection of SARS-CoV-2 from patient samples with high sensitivity and specificity. We obtained a limit of detection of 16 copies/μL with high specificity and at an affordable cost. The diagnostic test readout can be done with a real-time PCR thermocycler or with the naked eye in a blue-light transilluminator. Our method has been evaluated on a small set of clinical samples with promising results.


2022 ◽  
Vol 12 ◽  
Author(s):  
Katharina Radakovics ◽  
Claire Battin ◽  
Judith Leitner ◽  
Sabine Geiselhart ◽  
Wolfgang Paster ◽  
...  

Toll-like receptors (TLRs) are primary pattern recognition receptors (PRRs), which recognize conserved microbial components. They play important roles in innate immunity but also in the initiation of adaptive immune responses. Impurities containing TLR ligands are a frequent problem in research but also for the production of therapeutics since TLR ligands can exert strong immunomodulatory properties even in minute amounts. Consequently, there is a need for sensitive tools to detect TLR ligands with high sensitivity and specificity. Here we describe the development of a platform based on a highly sensitive NF-κB::eGFP reporter Jurkat JE6-1 T cell line for the detection of TLR ligands. Ectopic expression of TLRs and their coreceptors and CRISPR/Cas9-mediated deletion of endogenously expressed TLRs was deployed to generate reporter cell lines selectively expressing functional human TLR2/1, TLR2/6, TLR4 or TLR5 complexes. Using well-defined agonists for the respective TLR complexes we could demonstrate high specificity and sensitivity of the individual reporter lines. The limit of detection for LPS was below 1 pg/mL and ligands for TLR2/1 (Pam3CSK4), TLR2/6 (Fsl-1) and TLR5 (flagellin) were detected at concentrations as low as 1.0 ng/mL, 0.2 ng/mL and 10 pg/mL, respectively. We showed that the JE6-1 TLR reporter cells have the utility to characterize different commercially available TLR ligands as well as more complex samples like bacterially expressed proteins or allergen extracts. Impurities in preparations of microbial compounds as well as the lack of specificity of detection systems can lead to erroneous results and currently there is no consensus regarding the involvement of TLRs in the recognition of several molecules with proposed immunostimulatory functions. This reporter system represents a highly suitable tool for the definition of structural requirements for agonists of distinct TLR complexes.


Molecules ◽  
2021 ◽  
Vol 26 (20) ◽  
pp. 6163
Author(s):  
Aree Choodum ◽  
Nareumon Lamthornkit ◽  
Chanita Boonkanon ◽  
Tarawee Taweekarn ◽  
Kharittha Phatthanawiwat ◽  
...  

Benzo(a)pyrene (BaP) has been recognized as a marker for the detection of carcinogenic polycyclic aromatic hydrocarbons. In this work, a novel monolithic solid-phase extraction (SPE) sorbent based on graphene oxide nanoparticles (GO) in starch-based cryogel composite (GO-Cry) was successfully prepared for BaP analysis. Rice flour and tapioca starch (gel precursors) were gelatinized in limewater (cross-linker) under alkaline conditions before addition of GO (filler) that can increase the ability to extract BaP up to 2.6-fold. BaP analysis had a linear range of 10 to 1000 µgL−1 with good linearity (R2 = 0.9971) and high sensitivity (4.1 ± 0.1 a.u./(µgL−1)). The limit of detection and limit of quantification were 4.21 ± 0.06 and 14.04 ± 0.19 µgL−1, respectively, with excellent precision (0.17 to 2.45%RSD). The accuracy in terms of recovery from spiked samples was in the range of 84 to 110% with no significant difference to a C18 cartridge. GO-Cry can be reproducibly prepared with 2.8%RSD from 4 lots and can be reused at least 10 times, which not only helps reduce the analysis costs (~0.41USD per analysis), but also reduces the resultant waste to the environment.


2006 ◽  
Vol 69 (3) ◽  
pp. 682-685 ◽  
Author(s):  
ORGUN DEVECİ ◽  
EMEL SEZGİN

In this study, skim milk powder was produced from cow's milk contaminated artificially with aflatoxin M1 (AFM1) at two different levels, 1.5 and 3.5 μg/liter (ppb), and the effects of process stages on the AFM1 contents were investigated. Pasteurization, concentration, and spray drying caused losses of about 16, 40, and 68%, respectively, in AFM1 content of the milk contaminated with 1.5 μg/liter AFM1, and losses of 12, 35, and 59%, respectively, in the milk contaminated with 3.5 μg/liter AFM1. These losses were found to be statisticially significant at the level of P < 0.01. After 3- and 6-month storage periods, AFM1 content of the skim milk powder produced from milk with 1.5 μg/liter AFM1 decreased by 2 and 5%, respectively, whereas these rates were 2 and 4%, respectively, for the skim milk powders made from milk with 3.5 μg/liter AFM1 (after adjustment for sample weight). Changes in AFM1 content of milk powder samples were found statistically insignificant (P > 0.05 and P > 0.01) for 3- and 6-month storage periods.


2020 ◽  
Vol 20 (5) ◽  
pp. 3148-3156 ◽  
Author(s):  
S. Nehru ◽  
Subramanian Sakthinathan ◽  
P. Tamizhdurai ◽  
Te-Wei Chiu ◽  
K. Shanthi

In the present work, a reduced graphene oxide and multiwalled carbon nanotube (RGO/MWCNTFe3O4) composite decorated with Fe3O4 magnetic nanoparticles was prepared as an electrochemical sensor. The surface morphology of the prepared composite was identified by scanning electron microscopy and X-ray diffraction. The electrochemical properties of the GCE/RGO/MWCNT-Fe3O4 electrode were investigated by electrochemical impedance spectroscopy, cyclic voltammetry and amperometry. The GCE/RGO/MWCNT-Fe3O4 electrode exhibited higher electrocatalytic performance towards the oxidation of hydrazine. In the optimal conditions, the GCE/RGO/MWCNT-Fe3O4 electrode showed a wide linear range (0.15–220 μM), low limit of detection (LOD) (0.75 μM), and high sensitivity (2.868 μA μM−1 cm−2). The prepared GCE/RGO/MWCNT-Fe3O4 electrode also had excellent repeatability, selectivity, and reproducibility. The practical application of the electrode was confirmed with various spiked water samples and demonstrated acceptable recovery.


2019 ◽  
Vol 57 (5) ◽  
Author(s):  
Padmapriya P. Banada ◽  
Srinidhi Deshpande ◽  
Sukalyani Banik ◽  
Darshini Shah ◽  
Ranie Koshy ◽  
...  

ABSTRACT Francisella tularensis, Bacillus anthracis, and Yersinia pestis are tier 1 select agents with the potential to rapidly cause severe disease. Rapid detection of these bacteria from patient samples at the point of care could contribute to improved clinical outcomes in the event of a bioterrorism attack. A multiplex nested PCR assay for detection of F. tularensis, B. anthracis, and Y. pestis directly from patient blood samples was developed using the GeneXpert system. The multiplex GeneXpert cartridge-based assay includes all necessary sample processing and amplification reagents. Blood samples spiked with different numbers of CFU were used to measure the analytical limit of detection (LOD) and dynamic range. Sensitivity was determined by testing spiked blood samples and negative-control blood in a blind manner. Specificity was determined by testing against nontarget pathogens and blood samples from clinical patients. The assay LOD was 8.5 CFU/ml for F. tularensis, 10 CFU/ml for B. anthracis, and 4.5 CFU/ml for Y. pestis. The sensitivity was 100% at the LOD for all three select agent bacteria in spiked patient blood samples. The assay specificity was 100% when it was tested against both nontarget pathogens and clinical patient blood samples. The total assay time was approximately 100 min. This automated assay, which is suitable for use at the point of care, identifies three select agents directly in blood without the need for enrichment with a high sensitivity within 100 min. This assay may enable rapid detection and treatment of patients infected with the target organisms in the event of a bioterrorism attack.


Chemosensors ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 58
Author(s):  
Pranlekha Traiwatcharanon ◽  
Wilai Siriwatcharapiboon ◽  
Chatchawal Wongchoosuk

High sodium ion (Na+) consumption leads to high blood pressure which causes many health issues. Real-time determination of Na+ content in food is still important to limit Na+ intake and control the taste of food. In this work, we have developed an electrochemical sensor based on agglomeration of silver nanoparticles (AgNPs) and graphene oxide (GO) modified on a screen-printed silver electrode (SPE) for Na+ detection at room temperature by using cyclic voltammetry (CV). The AgNPs were synthesized through a simple green route using Pistia stratiotes extract as a reducing agent under blue light illumination and mixed with the GO to be a Na+ selective sensing nanocomposite. The AgNPs/GO/SPE sensor showed high sensitivity (0.269 mA/mM/cm2), high selectivity, linear relationship (0–100 mM), good stability, and excellent reproducibility to Na+ detection as well as low limit of detection (9.344 mM) for food application. The interfering species such as K+, Zn2+, Na+, Mg2+, glucose, and ascorbic acid did not have any influence on the Na+ determination. The AgNPs/GO/SPE sensor was successfully applied to determine Na+ in real samples such as fish sauce and seasoning powder of instant noodle.


Sign in / Sign up

Export Citation Format

Share Document