RESUMEN
Strategies for rapid, effective nucleic acid processing hold tremendous significance to the clinical analysis of circulating tumor DNA (ctDNA), a family of important markers indicating tumorigenesis and metastasis. However, traditional techniques remain challenging to achieve efficient DNA enrichment, further bringing about complicated operation and limited detection sensitivity. Here, we developed an ion concentration polarization microplatform that enabled highly rapid, efficient enrichment and purification of ctDNA from a variety of clinical samples, including serum, urine, and feces. The platform demonstrated efficiently separating and enriching ctDNA within 30 s, with a 100-fold improvement over traditional methods. Integrating an on-chip isothermal amplification module, the platform further achieved 100-fold enhanced sensitivity in ctDNA detection, which significantly eliminated false-negative results in the serum or urine samples due to the low abundance of ctDNA. Such a simple-designed platform offers a user-friendly yet powerful diagnosis technique with a wide applicability, ranging from early tumor diagnosis to infection screening.
Asunto(s)
ADN Tumoral Circulante , Neoplasias , Ácidos Nucleicos , Humanos , Neoplasias/diagnóstico , Neoplasias/genética , ADN Tumoral Circulante/genética , Carcinogénesis , Técnicas de Amplificación de Ácido Nucleico/métodosRESUMEN
The aim of this paper is to design and numerically simulate the mass-transfer compartment and piezoelectric micropump of an implantable integrated microfluidic device for regular microdialysis-based nonenzymatic measurement of glucose level in diabetic patients. The device function is based on the process that the piezoelectric micropump pumps the dialysis fluid into the mass-transfer compartment microchannels, where the interstitial fluid (ISF) glucose diffusion into this dialysis fluid gives it a glucose content, then detected and measured in the sensor section. This diffusion takes place through the semipermeable membranes located in the microchannels at the base of the hollow microneedles entering the body skin painlessly. The value of dialysis fluid flow rate (1 µL/min) was chosen so that the best achievable recovery factor can be obtained while the size and time delay of system were being kept at the best minimum possible. In the mass-transfer compartment, the number of microneedles, the dimensions of microchannels and the thickness of membranes were selected so as to achieve the best appropriate recovery factor, minimum possible size as well as considering the fabrication feasibility. Furthermore, in the different parts of micropump, the materials and dimensions were chosen so as to provide the needed flow rate with the best minimum voltage, sufficiently small size and fabrication feasibility.
RESUMEN
Cytokines are important factors in the early diagnosis of autoimmune diseases and require high sensitivity, high selectivity and quantitative detection. We proposed a miniaturized electrochemical magneto-immunosensor (EC-MIS) on portable interleukin-6 (IL-6) detection based on this requirement. Firstly, a micro-fabricated working electrode is electrochemically modified with a hybrid of reduced graphene oxide (rGO) and gold nanoparticles (AuNPs). Increased surface area and enhanced charge transfer rate improve the performance of this immunosensor on sensitivity. Secondly, magnetic beads attached with the capture antibody (cAb) are employed in sandwich immunoassay. This kind of immunoassay is immobilized on the working electrode surface by an external magnet to enrich the analyte IL-6. Thirdly, the last two features are combined and integrated on a microfluidic device in order to restrict the sample at certain areas and ease the operation of detection. With our prototypic EC-MIS operated in amperometric mode, we have achieved the detection of IL-6 with a linear range from 0.97 to 250 pg/mL and a limit of detection (LOD) of 0.42 pg/mL. Real serum samples were demonstrated and compared with benchtop equipment's results.