Simultaneous detection of ovarian cancer related miRNA biomarkers with carboxylated graphene oxide modified electrochemical biosensor platform.

Ovarian cancer, known as "silent killer", is a gynocological cancer with high mortality that usually diagnosed in the late stages. Gold standard immunoassay technique is evaluation of CA-125 levels which is not merely specific to ovarian cancer. Therefore, there is a need for sensitive determination of more specific biomarkers. miR-200 family is RNA nucleic acids that known to be upregulated in the presence of ovarian cancer. Since diagnosis based on a single biomarker is prone to generate misleading results, it is important to develop point-of-care systems that allow diagnosis of multiple miRNAs. Herein, an electrochemical nanobiosensor platform was developed for the multiplexed and simultaneous detection of miR-200c and miR-141. Biorecognition part was constitutued of methylene blue and ferrocene labeled hairpin DNA probes immobilized onto carboxylated graphene oxide modified pencil graphite electrodes. Their hybridization with miRNAs were examined upon "signal-off" approach using Square Wave Voltammetry. The platform demonstrated a linear detection range of 0.1 pM to 10 nM for both miR-141 and miR-200c, with low detection limits of 0.029 pM and 0.026 pM, respectively. We assume that the developed biosensor platform may pave the way in early diagnosis of the disease and the development of more effective treatment strategies.

Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor.

The CRISPR-Cas9 system has facilitated the genetic modification of various model organisms and cell lines. The outcomes of any CRISPR-Cas9 assay should be investigated to ensure/improve the precision of genome engineering. In this study, carbon nanotube-modified disposable pencil graphite electrodes (CNT/PGEs) were used to develop a label-free electrochemical nanogenosensor for the detection of point mutations generated in the genome by using the CRISPR-Cas9 system. Carbodiimide chemistry was used to immobilize the 5'-aminohexyl-linked inosine-substituted probe on the surface of the sensor. After hybridization between the target sequence and probe at the sensor surface, guanine oxidation signals were monitored using differential pulse voltammetry (DPV). Optimization of the sensitivity of the nanogenoassay resulted in a lower detection limit of 213.7 nM. The nanogenosensor was highly specific for the detection of the precisely edited DNA sequence. This method allows for a rapid and easy investigation of the products of CRISPR-based gene editing and can be further developed to an array system for multiplex detection of different-gene editing outcomes.

Aptamer-based electrochemical biosensing strategy toward human non-small cell lung cancer using polyacrylonitrile/polypyrrole nanofibers.

In the present study, a sensitive electrochemical aptamer-based biosensing strategy for human non-small cell lung cancer (NSCLC) detection was proposed using nanofiber-modified disposable pencil graphite electrodes (PGEs). The composite nanofiber was comprised of polyacrylonitrile (PAN) and polypyrrole (PPy) polymers, and fabrication of the nanofibers was accomplished using electrospinning process onto PGEs. Development of the nanofibers was confirmed using scanning electron microscopy (SEM). The high-affinity 5'-aminohexyl-linked aptamer was immobilized onto a PAN/PPy composite nanofiber-modified sensor surface via covalent bonding strategy. After incubation with NSCLC living cells (A549 cell line) at 37.5 °C, the recognition between aptamer and target cells was monitored by electrochemical impedance spectroscopy (EIS). The selectivity of the aptasensor was evaluated using nonspecific human cervical cancer cells (HeLa) and a nonspecific aptamer sequence. The proposed electrochemical aptasensor showed high sensitivity toward A549 cells with a detection limit of 1.2 × 103 cells/mL. The results indicate that our label-free electrochemical aptasensor has great potential in the design of aptasensors for the diagnostics of other types of cancer cells with broad detection capability in clinical analysis. Graphical abstract.