Centrifugal Microfluidic Cell Culture Platform for Physiologically Relevant Virus Infection Studies: A Case Study with HSV-1 Infection of Periodontal Cells.

Static well plates remain the gold standard to study viral infections in vitro, but they cannot accurately mimic dynamic viral infections as they occur in the human body. Therefore, we established a dynamic cell culture platform, based on centrifugal microfluidics, to study viral infections in perfusion. To do so, we used human primary periodontal dental ligament (PDL) cells and herpes simplex virus-1 (HSV-1) as a case study.

Development of nucleocapsid-specific monoclonal antibodies for SARS-CoV-2 and their ELISA diagnostics on an automatic microfluidic device.

Coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has threatened public health globally, and the emergence of viral variants has exacerbated an already precarious situation. To prevent further spread of the virus and determine government action required for virus control, accurate and rapid immunoassays for SARS-CoV-2 diagnosis are urgently needed. In this study, we generated monoclonal antibodies (mAbs) against the SARS-CoV-2 nucleocapsid protein (NP), compared their reactivity using an enzyme-linked immunosorbent assay (ELISA), and selected four mAbs designated 1G6, 3E10, 3F10, and 5B6 which have higher reactivity to NP and viral lysates of SARS-CoV-2 than other mAbs.

Prediction of acute rejection in kidney transplanted patients based on the point-of-care isothermal molecular diagnostics platform.

In this study, we proposed an advanced point-of-care molecular diagnostic technology to evaluate the acute rejection (AR) in kidney transplanted patients. On the contrary to the conventional PCR method, we developed a colorimetric loop mediated isothermal amplification (LAMP) for quantitative analysis of the six biomarkers related to AR (CD3ϵ, IP-10, Tim-3-HAVCR2, CXCL9, PSMB9, C1QB) with a reference gene (18S rRNA). Using urinary cDNA samples of transplanted patients, it turned out that three biomarkers among six, namely IP-10, Tim-3-HAVCR2 and C1QB, have significant discrepancy in quantity between the stable graft (STA) patient and the AR patient.

A portable centrifugal genetic analyzer for multiplex detection of feline upper respiratory tract disease pathogens.

We present a portable genetic analyzer with an integrated centrifugal disc which is equipped with a glass-filter extraction column for purifying nucleic acid (NA) and multiple reaction chambers for analyzing major feline upper respiratory tract disease (FURTD) pathogens. We targeted four kinds of FURTD including Feline herpesvirus 1 (FHV), Mycoplasma felis (MPF), Bordetella bronchiseptica (BDB), and Chlamydophila felis (CDF). The portable genetic analyzer consists of a spinning motor, two pairs of Peltier heaters, two Minco heater, fluorescent optics, a touchscreen, and software for data analysis, so loop-mediated isothermal amplification (LAMP) or polymerase chain reaction (PCR) can be performed.

High-throughput human DNA purification on a centrifugal microfluidic device for rapid forensic sex-typing.

We introduce a novel centrifugal disc for purifying nucleic acid (NA) in a high-throughput manner to perform the human sex-typing of forensic samples. The centrifugal disc was designed with double-sided etched channels to fabricate 30 extraction units, which was capable of purifying 30 forensic samples in a single run. In order to introduce the washing solution (70% ethanol) and the elution buffer for the 30 extraction units in an automatic manner, we designed the aliquoting chambers that were connected with a zigzag delivery channel.

Quantification of colorimetric isothermal amplification on the smartphone and its open-source app for point-of-care pathogen detection.

The increasing risk of infectious pathogens, especially in the under-developed countries, is demanding the development of point-of-care (POC) nucleic acid testing in the low-resource setting conditions. Here, we describe a methodology for colorimetric quantitative analysis of nucleic acid using an easy-to-build smartphone-based platform, offering low-cost, portability, simplicity in operation, and user-friendliness. The whole system consists of a hand-held box equipped with a smartphone, a film heater, a white LED, a loop-mediated isothermal amplification (LAMP) chip, and a DC converter, and all the processes were powered by a portable battery of 5 V.

An Integrated Smartphone-Based Genetic Analyzer for Qualitative and Quantitative Pathogen Detection.

The use of the smartphone is an ideal platform to realize the future point-of-care (POC) diagnostic system. Herein, we propose an integrated smartphone-based genetic analyzer. It consists of a smartphone and an ntegrated tic analysis unit (i-Gene), in which the power of the smartphone was utilized for heating the gene amplification reaction, and the camera function was used for imaging the colorimetric change of the reaction for quantitative and multiplex foodborne pathogens.

Centrifugal microfluidic device for the high-throughput synthesis of Pd@AuPt core-shell nanoparticles to evaluate the performance of hydrogen peroxide generation.

We propose a novel high-throughput screening platform using a centrifugal microfluidic device for producing combinatorial tri-metallic catalysts. The centrifugal device was designed to perform 60 reactions under different conditions on a single device. As a model to search for an optimal tri-metallic catalyst, we synthesized a variety of Pd@AuPt nanoparticles (NPs), in which Pd nanocubes served as a core, and Au and Pt atoms formed a shell.

Nucleic acid diagnostics on the total integrated lab-on-a-disc for point-of-care testing.

Since the emergence of the lab-on-a-chip technology in 1979, a variety of microfluidic devices have been developed and utilized for chemical and biological applications. Among the microfluidic devices, the centrifugal microfluidic device or lab-on-a-disc (LOAD) has advanced remarkably due to simple operation by the rotation, total integration, and high-throughput capability. Moreover, the centrifugal microdevices do not need complex tubing and pumping systems, which render them ideal for point-of-care testing (POCT) system.

Point-of-care genetic analysis for multiplex pathogenic bacteria on a fully integrated centrifugal microdevice with a large-volume sample.

We present a fully integrated portable centrifugal microsystem for multiplex detection of food poisoning bacteria with a large volume of sample up to 1 mL. The microsystem consists of a portable genetic analyzer and a fully integrated centrifugal microdevice. The centrifugal microdevice is designed with two units: a 3D printed solution-loading cartridge and a centrifugal microfluidic disc.