RESUMO
An integrated electrochemical immunoassay is described for the determination of circulating tumor cells (CTCs). For the first time, Ketjen black (KB), which is a superconductive carbon material, was incorporated with Au nanoparticles (AuNPs) and used to modify the surface of gold electrodes. A cocktail of anti-epithelial cell adhesion molecules (EpCAM) and anti-vimentin antibodies was chosen to capture the CTCs. Palladium-iridium-boron-phosphorus alloy-modified mesoporous nanospheres (PdIrBPMNS) served as a catalytic tag to amplify the current signal. Glycine-HCl (Gly-HCl) was used as an antibody eluent to release and collect the captured CTCs from the electrodes for further clinical research without compromising cell viability. The response of the method increased linearly from 10 to 1 × 106 cells mL-1 CTCs, while the detection limit was calculated to be as low as 2 cells mL-1. This method was successfully used to determine CTCs in spiked blood samples and demonstrated good recovery. Graphical abstractKetjen black/AuNPs was incorporated in the electrochemical platform to enhance the electron transfer ability of the electrode surface. PdIrBP mesoporous nanospheres were used to amplify DPV signal in this assay. The introduction of Gly-HCl realized nondestructive recovery of circulating tumor cells.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Nanosferas/química , Células Neoplásicas Circulantes/patologia , Fuligem/química , Boro/química , Condutividade Elétrica , Humanos , Irídio/química , Paládio/química , Tamanho da Partícula , Fósforo/química , Porosidade , Propriedades de Superfície , Células Tumorais CultivadasRESUMO
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.
Assuntos
Materiais Biomiméticos/química , Colorimetria/métodos , Dendrímeros/química , Proteínas de Fusão bcr-abl/genética , Ouro/química , Peroxidase/metabolismo , Platina/química , Nanopartículas Metálicas/química , Nanocompostos/química , Reação em Cadeia da PolimeraseRESUMO
The study aims to assess the accuracy of the arterial blood gas (ABG) analysis in measuring hemoglobin, potassium, sodium, and glucose concentrations in comparison to standard venous blood analysis among patients diagnosed with chronic obstructive pulmonary disease (COPD). From January to March 2023, results of ABG analysis and simultaneous venous blood sampling among patients with COPD were retrospectively compared, without any intervention being applied between the two methods. The differences in hemoglobin, potassium, sodium, and glucose concentrations were assessed using a statistical software program (R software). There were significant differences in the mean concentrations of hemoglobin (p < 0.001), potassium (p < 0.001), and sodium (p = 0.001) between the results from ABG and standard venous blood analysis. However, the magnitude of the difference was within the total error allowance (TEa) of the United States of Clinical Laboratory Improvement Amendments (US-CLIA). As for the innovatively studied glucose concentrations, a statistically significant difference between the results obtained from ABG (7.8 ± 3.00) mmol·L-1 and venous blood (6.72 ± 2.44) mmol·L-1 was noted (p < 0.001), with the difference exceeding the TEa of US-CLIA. A linear relationship between venous blood glucose and ABG was obtained: venous blood glucose (mmol·L-1) = - 0.487 + 0.923 × ABG glucose (mmol·L-1), with R2 of 0.882. The hemoglobin, potassium, and sodium concentrations in ABG were reliable for guiding treatment in managing COPD emergencies. However, the ABG analysis of glucose was significantly higher as compared to venous blood glucose, and there was a positive correlation between the two methods. Thus, a linear regression equation in this study combined with ABG analysis could be helpful in quickly estimating venous blood glucose during COPD emergency treatment before the standard venous blood glucose was available from the medical laboratory.
Assuntos
Doença Pulmonar Obstrutiva Crônica , Sódio , Humanos , Glucose , Glicemia , Potássio , Estudos Retrospectivos , Gasometria/métodos , Hemoglobinas/análiseRESUMO
Accurate detection of circulating tumor cells (CTCs) has a pivotal role in the metastasis monitoring and prognosis of tumor. In this work, an ultrasensitive electrochemical cytosensor was developed based on excellent electrocatalytic materials and a dual recognition strategy. Herein, novel branched PtAuRh trimetallic nanospheres (b-PtAuRh TNS) were synthesized for the first time by a facile one-pot method, which had a huge specific surface area and outstanding catalytic activity. B-PtAuRh TNS linked with aptamers targeting mucin1 (MUC1) were served as signal tags to amplify the signal. As electrode modified material, the nanocomposites of Cabot carbon black (BP2000) and AuNPs were used to improve the electron transfer efficiency of electrode. In addition to using b-PtAuRh TNS labeled anti-MUC1 aptamers as signal probes, anti-EpCAM antibodies were worked as capture probes to achieve dual recognition of target cells. In other words, only cells expressing both MUC1 and EpCAM could produce electrochemical signal. The constructed cytosensor presented a wide linear range (5 - 1 × 106 cells mL-1) and a low detection limit (1 cell mL-1). It was worth noting that the proposed cytosensor could detect CTCs in clinical blood samples. To sum up, the developed cytosensor might become a promising detection platform for cancer diagnosis and tumor metastasis.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Nanosferas , Células Neoplásicas Circulantes , Técnicas Eletroquímicas , Ouro , Humanos , Limite de DetecçãoRESUMO
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.