A highly sensitive fluorescent immunosensor for sensitive detection of nuclear matrix protein 22 as biomarker for early stage diagnosis of bladder cancer.

A novel strategy is reported for highly sensitive, rapid, and selective detection of nuclear matrix protein NMP22 using two-color quantum dots based on fluorescence resonance energy transfer (FRET). Quantum dots (QDs) are highly advantageous for biological imaging and analysis, particularly when combined with (FRET) properties of semiconductor quantum dot (QDs) are ideal for biological analysis to improve sensitivity and accuracy. In this FRET system narrowly dispersed green emitting quantum dot CdTe core is used as a donor and labelled by monoclonal (mAb) antibody, while orange emitting quantum dot CdTe/CdS core shell is used as an accepter and labelled by polyclonal (pAb) antibody. The quantum dots are labelled by antibodies using EDC/NHS as crosslinking agent. Bovine serum albumin (BSA) solution was added to block nonspecific binding sites. The fluorescence intensity of QDs accepter decreased linearly with the increasing concentrations of NMP22 from 2-22 pg mL-1 due to FRET system and fluoroimmunoassay reaction. This method has good regression coefficient (R 2 = 0.998) and detection limit was 0.05 pg mL-1. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications.

Paper based colorimetric detection of miRNA-21 using Ag/Pt nanoclusters.

Abnormal expression of MicroRNA-21 (miRNA-21) is considered to be a reliable biomarker for the early diagnosis of cancer. In this work, a novel paper based biosensor was fabricated to detect sub-micro molar concentrations of miRNA-21 based on peroxidase mimetic activity of DNA-templated Ag/Pt nanoclusters (DNA-Ag/Pt NCs), which could catalyze the reaction of hydrogen peroxide and 3,3',5,5' tetramethylbenzidine (TMB), to produce a blue color. The Mechanism of reaction was based on the inhibition effect of miRNA-21 on peroxidase-like activity of nanosensor which resulted to quantitative determination of miRNA-21 concentration. It was found that miRNA-21 could be linearly detected in the range from 1-700 pM (A652 = 0.16x-0.96, R2 = 0.99; x = -log [miRNA-21]) with a detection limit of 0.6 pM. Moreover, a paper assay was carried out on a Y-shaped paper-based microfluidic device in order to use the distinctive features of micro-channels such as short response time, very low reagent volume, low fabrication cost, etc. After performing paper based assay, a good linear range was observed between 10-1000 pM (y = 0.06x+147.48, R2 = 0.99; x = [miRNA-21]) with detection limit of 4.1 pM. The practical application of proposed method for detection of miRNA-21 in real sample was assayed in the human urine sample and indicated the colorimetric method had acceptable accuracy.

Enhanced peroxidase-like activity of platinum nanoparticles decorated on nickel- and nitrogen-doped graphene nanotubes: colorimetric detection of glucose.

A nanostructured catalyst is introduced that demonstrates peroxidase mimicking activity. It consists of nickel- and nitrogen-doped graphene nanotubes loaded with platinum nanoparticles. Pt-decorated Ni-doped nitrogen-rich graphitic nanotube (Pt/Ni@NGT) was synthesized using a two-step procedure in which the precursors were first refluxed to form a supramolecular assembly followed by a pyrolysis and leaching step to form nanotubes. Afterwards, Pt was decorated on the outer surface of nanotube by an ultrasound assisted method. Pt/Ni@NGT was characterized by XPS, TEM, SEM, and HAADF-STEM. The as-prepared Pt/Ni@NGT nanostructure was used for the detection of glucose via catalyzing the oxidation of a substrate, 3,3',5,5'-tetramethylbenzidine (TMB), to form a blue product (ox-TMB), thereby enabling colorimetric assay for enzymatically generated H2O2. The nanostructure exhibited excellent biocompatibility and led to highly efficient immobilization and retention of GOx. The method has a linear response in the 43 pM to 220 µM glucose concentration range, a detection limit as low as 1 pM and a limit of quantification of 3.4pM, along with good reproducibility(< 3%). A paper based visual microfluidic assay was also worked out that has an analytical range that extends from 0.1-50 mM. It is simple and rapid enough to be useful as a glucose home test.. The method was successfully applied to the determination of glucose in tear and saliva samples. Graphical abstract Graphene nanotubes doped with nitrogen and nickel (Ni@NGT) have been synthesized as the support to construct the unique Pt/Ni@NGT for providing artificial peroxidase activity for the GOx-based detection of glucose, which was further used for the construction of a glucose paper assay.

A colorimetric paper sensor for citrate as biomarker for early stage detection of prostate cancer based on peroxidase-like activity of cysteine-capped gold nanoclusters.

Citrate is currently considered a preferred biomarker for the early stage detection of prostate cancer. In the present work, based on the highly efficient catalytic properties of gold nanoclusters, a novel system for optical determination of citrate was successfully established under optimized conditions. Cysteine-capped gold nanoclusters (Cys-AuNCs) are shown to have an intrinsic peroxidase-mimetic activity. In the presence of H2O2, Cys-AuNCs nanostructures are able to catalyse the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) with high efficiency to produce a blue dye (with an absorbance maximum at 650 nm). Citrate has carboxylic and hydroxyl groups that can bind with free amino and free carboxyl cysteine groups via hydrogen bonds, thus creating a coating on the surface of the gold nanocluster and inhibiting the cluster oxidation activity. Accordingly, a visual, sensitive and simple colorimetric method using Cys-AuNCs as peroxidase mimetic was developed for detecting citrate. A suitable linear relationship for citrate was obtained for the range of 0.5 to 1000 µM. The limit of detection (LOD) of the proposed method was calculated as 0.1 µM and the relative standard deviation (RSD) was obtained to be less than 4.0%. Moreover, the biosensor was used to perform a paper assay on a Y-shaped microfluidic device and make use of the distinctive features of microchannels such as short response time, very low reagent volume required, low fabrication cost etc. A detection limit of 0.4 µM was achieved through the paper test and a good linear range was observed between 1.0 µM-10 mM. The proposed method was further applied to citrate detection in the human urine sample.