Colorimetric determination of BCR/ABL fusion genes using a nanocomposite consisting of Au@Pt nanoparticles covered with a PAMAM dendrimer and acting as a peroxidase mimic.

A colorimetric assay is described for the detection of BCR/ABL fusion genes. Polyamidoamine (PAMAM) dendrimers were placed on peroxidase (POx) mimicking Au@Pt nanoparticles to form a nanocomposite of type Au@Pt-PAMAM. Capture DNA probe is a designed nucleic acid strand that specifically binds target DNA to the surface of the electrode. The capture probe was attached to magnetic beads via biotin and avidin interaction. The hairpin structure of the capture probe can only be opened by the complementary BCR/ABL DNA. This results in a highly specific assay. The POx-mimicking property of the Au@Pt-PAMAM causes the formation of a blue dye by reaction of H2O2 and 3,3,3',3'-tetramethylbenzidine (TMB) which is measured by a microplate reader. Under optimum conditions, the absorbance increases linearly the 1 pM to 100 nM BCR/ABL concentration range, and the detection limit is as low as 190 fM. The method is highly selective and was successfully applied to the determination of fusion genes in spiked real samples. Conceivably, it possesses a large potential in clinical testing of patients suffering from chronic myeloid leukemia. Graphical abstract Au@PtNP, an efficient catalyst, is bound with polyamidoamine (PAMAM) dendrimer to amplify the colorimetric signal. With the introduction of streptavidin-magnetic beads to remove non-specific signals, a novel colorimetric sensor is constructed to detect BCR/ABL fusion genes.

Double signal enhancement strategy based on rolling circle amplification and photoinduced electron transfer for ultrasensitive fluorometric detection of methylated DNA.

The authors describe a novel assay for the detection of methylated DNA site. Rolling circle amplification and CdSe/ZnS quantum dots with high fluorescence efficiency are applied in this method. The CdSe/ZnS quantum dots act as electron donors, and hemin and oxygen (derived from hydrogen peroxide act as acceptors in photoinduced electron transfer. The assay, best performed at excitation/emission peaks of 450/620 nm, is sensitive and specific. Fluorometric response is linear in the 1 pM to 100 nM DNA concentration range, and the lowest detectable concentration of methylated DNA is 142 fM (S/N = 3). The method is capable of recognizing 0.01% methylated DNA in a mixture of methylated/unmethylated DNA. Graphical abstract A novel method for methylated sites detection in DNA is established. Rolling circle amplification and photoinduced electron transfer. CdSe/ZnS quantum dots with high fluorescence efficiency act as the electron donor, while G-quadruplex/hemin and hydrogen peroxide derived oxygen act as electron acceptor. It presents a linear response towards 1 pM to 100 nM methylated DNA with a correlation coefficient of 0.9968, and the lowest detectable concentration of methylated DNA was 142 fM, with selectivity significantly superior to other methods.

A novel cytosensor based on Pt@Ag nanoflowers and AuNPs/Acetylene black for ultrasensitive and highly specific detection of Circulating Tumor Cells.

Circulating tumor cells (CTCs), as the cellular origin of metastasis, are cancer cells that break away from a primary tumor and circulate in the peripheral blood. And they provide a wealth of information about tumor phenotype. Here, this work reported a novel ultrasensitive immunoassay protocol for the detection of CTCs by using Pt@Ag nanoflowers (Pt@AgNFs) and AuNPs/Acetylene black (AuNPs/AB) nanomaterial. In the established approach, AuNPs/AB nanomaterial was used as substrate material to increase the specific surface area and enhance the conductivity of the gold electrode. Protein G was used for oriented immobilization of capture antibody, which strongly improved the capture efficiency of MCF-7 cells. The innovatively synthesized Pt@AgNFs by our group with high specific surface area and good biocompatibility were not only as the carriers of signal antibodies (Ab2) but also catalyzed the reduction of H2O2, which effectually amplified the current signal. A linear relationship between current signals and the concentrations of CTCs was obtained in the range from 20 to 1×106 cells mL-1 and the detection limit is as low as 3 cells mL-1 on condition of acceptable stability and reproducibility. Furthermore, the as-proposed cytosensor showed excellent performance in the detection of CTCs in human blood samples. These results suggest that the proposed cytosensor will be a promising application for accurately quantitative detection of CTCs.