Electrochemical nano-genosensor for highly sensitive detection of miR-21 biomarker based on SWCNT-grafted dendritic Au nanostructure for early detection of prostate cancer.

MicroRNAs (miRNAs) appear as a novel reliable candidate in biomarkers for early diagnosis of cancer. Due to their roles in various types of cancer, their potential as a diagnostic biomarker is getting more attention. Here, a novel electrochemical biosensor for detection of miR-21 was demonstrated, through combining the advantages of electrochemical methods and nanomaterials with the selectivity of oligonucleotides, based on thiolated receptor probe-functionalized dendritic gold nanostructures (den-Au) via the self-assembly monolayer (SAM) process which grafted on the single-wall carbon nanotubes (SWCNTs) platform on the surface of the fluorine-doped tin oxide (FTO) electrode. Cadmium ions (Cd2+) were used as signal units and also signal amplification substance which labeled before on miR-21 target. The oxidation signal of Cd2+ as a signal unit was measured by differential pulse voltammetry (DPV) technique that had a very wide linear relationship with the concentration of miR-21 target (0.01 fmol L-1 to 1 µmol L-1) and low experimental detection limit of 0.01 fmol L-1. Furthermore, fabricated biosensor showed acceptable performance in human serum samples and also good selectivity indiscriminate between the complementary target and non-complementary one, so this nano-genosensor can clinically be used for prostate cancer diagnosis through the detection of miR-21 in human serum samples.

An electrochemical paper based nano-genosensor modified with reduced graphene oxide-gold nanostructure for determination of glycated hemoglobin in blood.

Hemoglobin A1c (HbA1c) is a standard biomarker to measure long-term average glucose concentration for diagnosis and monitoring of diabetes. Various methods have been reported for measuring HbA1c, however, portable and precise determination is still challenging. Herein, a new highly sensitive electrochemical nanobiosensor is developed for the specific determination of HbA1c. A nanocomposite of reduced graphene oxide (rGO) and gold with hierarchical architecture structure was electrochemically deposited on a cheap and flexible graphite sheet (GS) electrode. The nanocomposite increased the surface area, improved the electron transfer on the electrode surface and augmented the signal. It also provided a suitable substrate for linkage of thiolated DNA aptamer as a bioreceptor on the electrode surface by strong covalent bonding. The quantitative label free detection was carried out by differential pulse voltammetry (DPV) in a phosphate-buffered saline (PBS) solution containing redox probe Fe(CN)63-/4-. The detection is based on insulating the surface in presence of HbA1c and decreasing the current, which is directly related to the HbA1c concentration. The nanobiosensor demonstrated high sensitivity of 269.2 µA. cm-2, wide linear range of 1 nM-13.83 µM with a low detection limit of 1 nM. The biosensor was successfully used for measuring HbA1c in blood real sample. Furthermore, it is promising to use it as a part of a point of care device for low-invasive screening and management of diabetes.

Label-free ultrasensitive detection of breast cancer miRNA-21 biomarker employing electrochemical nano-genosensor based on sandwiched AgNPs in PANI and N-doped graphene.

MicroRNAs (miRNAs) are small, endogenous, noncoding RNAs, shown to be expressed abnormally in many tumors and identified as predictive biomarkers for early diagnosis of several cancers including the breast. Therefore, the label-free and highly sensitive detection of miRNAs is of critical significance. In this work, a highly sensitive and label-free nano-genosensor is developed for the detection of miRNA-21, a known breast cancer biomarker, based on a specific architecture of nitrogen-doped functionalized graphene (NFG), silver nanoparticles (AgNPs), and polyaniline (PANI) that resulted in a remarkable effect on signal amplification. Following the successful functionalization of the nanocomposite and immobilization of the specific sequence of the aminated complementary oligonucleotide of miRNA-21, the detection was performed using differential pulse voltammetry (DPV). The oxidation peak current of the redox probe under optimal conditions was determined to monitor the event hybridization of miRNA-21 biomarker. Applying this highly sensitive and optimized nano-biosensor enabled detection of a wide dynamic range of 10 fM-10 µM with a sensitivity of 2.5 µA cm-2 and a low detection limit of 0.2 fM. This nano-biosensor also demonstrated highly reproducible results in the analysis of blood samples, with recoveries between 94% and 107%, and could be used for early detection of breast cancer by direct detection of the miRNA-21 in real clinical samples without any need to sample preparation, RNA extraction and/or amplification.

Femtomolar level detection of RASSF1A tumor suppressor gene methylation by electrochemical nano-genosensor based on Fe3O4/TMC/Au nanocomposite and PT-modified electrode.

The alterations in DNA methylation pattern have been identified as one of the most frequent molecular phenomenon in human cancers. The RASSF1A tumor suppressor gene was shown to be often inactivated by hypermethylation of its promoter region. In the present study, a novel chip format sandwich electrochemical genosensor has been developed for the analysis of gene-specific methylation using Fe3O4/N-trimethyl chitosan/gold (Fe3O4/TMC/Au) nanocomposite as tracing tag to label DNA probe and polythiophene (PT) as immobilization platform of sensing element. However, no attempt has yet been made to conjugate DNA probe to Fe3O4/TMC/Au nanocomposite as electrochemical label for strip-based genosensing. Cyclic voltammetric (CV) analysis indicated that modification procedure was well performed. Differential pulse voltammetry (DPV) was employed for quantitative assessment of RASSF1A DNA promoter methylation. The electrochemical measurements accomplished using non-specific DNA fragments mixed with samples, revealed the high specificity and selectivity in methylation analysis by means of this DNA nanobiosensor. With the linear range of concentration from 1 × 10(-14)M to 5 × 10(-9)M and the detection limit of 2 × 10(-15)M, this new strategy has shown such a promising application to be used for universal analysis of any DNA sequence.