Integrated High-Throughput Centrifugal Microfluidic Chip Device for Pathogen Detection On-Site.

An integrated and high-throughput device for pathogen detection is crucial in point-of-care testing (POCT), especially for early diagnosis of infectious diseases and preventing the spread of infection. We developed an on-site testing platform that utilizes a centrifugal microfluidic chip and automated device to achieve high-throughput detection. The low-power (<32 W), portable (220 mm × 220 mm × 170 mm, 4 kg) device can complete bacterial lysis, nucleic acid extraction and purification, loop-mediated isothermal amplification (LAMP) reaction, and real-time fluorescence detection. Magnetic beads for nucleic acid adsorption can be mixed by applying electromagnetic fields and centrifugal forces, and the efficiency of nucleic acid extraction is improved by 60% compared to the no-mixing group. The automated nucleic acid extraction process achieves equivalent nucleic acid extraction efficiency in only 40% of the time consumed using the kit protocol. By designing the valve system and disc layout, the maximum speed required for the centrifugal microfluidic chip is reduced to 1500 rpm, greatly reducing the equipment power consumption and size. In detecting E. coli, our platform achieves a limit of detection (LOD) of 102 CFU/mL in 60 min. In summary, our active centrifugal microfluidic platform provides a solution for the integration of complex biological assays on turntables, with great potential in the application of point-of-care diagnosis.

High-throughput identification of meat ingredients in adulterated foods based on centrifugal integrated purification-CRISPR array.

Rapid, accurate, and sensitive analytical methods for the detection of food fraud are now an urgent requirement in the global food industry to ensure food quality. In response to this demand, a centrifugal integrated purification-CRISPR array for meat adulteration (CIPAM) was established. In detail, CIPAM system combines microneedles for DNA extraction and RAA-CRISPR/Cas12a integrated into a centrifugal microfluidic chip for the detection of meat adulteration. The RAA-CRISPR/Cas12a reaction reagents were pre-embedded into the different reaction chambers on the microfluidic chip to achieve the streamline of operations, markedly simplifying the detection process. The whole reaction was completed within 30 min with a detection limit of 0.1 % (w/w) in pig, chicken, duck, and lamb products. Referring to the results of the standard method, CIPAM system achieved 100 % accuracy. The automatic multiplex detection process implemented in the developed CIPAM system met the needs of food regulatory authorities.

Detection of VEGF165 in Whole Blood by Differential Pulse Voltammetry Based on a Centrifugal Microfluidic Chip.

For the rapid and sensitive detection of vascular endothelial growth factor 165 (VEGF165) in clinical blood samples, a microfluidic sensing chip that integrates a centrifugal separation pretreatment unit and a composite nanosensing film was proposed in this paper. An efficient sensing strategy and method was established. The blood sample was first separated and extracted by centrifugal force on the centrifugal microfluidic chip within 5 min after injection. The separated plasma can be automatically transferred through the designed microchannels to the detection area integrated electrodes for subsequent differential pulse voltammetric detection. The Au NPs/MCH/Apt2 sensing film was constructed on the surface of the Au working electrode. A sandwich sensing strategy based on "double aptamers" and "nanoprobe" for VEGF165 detection was established, by which the synthetic Apt1/PThi/Au NP nanoprobe was applied to capture VEGF165 in plasma and bind to the sensing film. By this method, the detection limit of VEGF165 in whole blood was 0.67 pg/mL and the linear range was between 1 pg and 10 ng, which met the needs of clinical VEGF165 detection. It was illustrated that the proposed methodology based on the centrifugal microfluidic chip had potential application prospects in the development of the point-of-care testing fields.

Multiwavelength Regression Algorithm for Eliminating Chamber Surface Effects of Microfluidic Chips.

An ultraviolet visible (UV-Vis) spectrophotometric and multiwavelength linear regression (MLR) method was developed for eliminating the influence of the surface quality of centrifugal microfluidic chips on the accuracy of their absorbance detection. The regression model is based on scalar scattering theory. The method was validated with cuvettes with different surface quality and Orange G (orange gelb) dye. The coefficients of variation (CVs) of the predicted solution concentration ratios in different cuvettes were < 1%, and the relative errors were < 1.5%. The model was shown to have higher accuracy and precision than that of traditional methods.

Design and preparation of centrifugal microfluidic chip integrated with SERS detection for rapid diagnostics.

A microfluidic SERS chip integrated with blood separation and in-situ detection was designed and fabricated for the rapid detection of clinical blood samples. Each functional unit in the microfluidic SERS chip consist of separation-decantation cavity based on centrifugal separation principle, mixing channels and SERS detection chamber built with integrated nano-Au on Ag film microstructure. The serum creatinine was selected as a typical sample to demonstrate the capability of microfluidic SERS chip. It was found that the creatinine SERS characteristic peaks at 678 cm-1 can be effectively identified and the detection limit could be as low as 4.42 × 10-3 µmol mL-1 in water. The blood samples were also tested in microfluidic SERS chip. The whole separation and test process could be completed within 2 min, which is a significant improvement in the field of creatinine detection. The whole blood of six cases clinical blood samples were also tested, and the results were consistent with the enzymatic results. The developed microfluidic SERS chip has advantages including reduction of the required quantity of blood sample, reusable and easy to operate. It is expected to provide a new method for rapid diagnostics.

Multitarget sensing of glucose and cholesterol based on Janus hydrogel microparticles.

A visualized sensing method for glucose and cholesterol was developed based on the hemispheres of the same Janus hydrogel microparticles. Single-phase and Janus hydrogel microparticles were both generated using a centrifugal microfluidic chip. For glucose sensing, concanavalin A and fluorescein labeled dextran used for competitive binding assay were encapsulated in alginate microparticles, and the fluorescence of the microparticles was positively correlated with glucose concentration. For cholesterol sensing, the microparticles embedded with γ-Fe2O3 nanoparticles were used as catalyst for the oxidation of 3,3',5,5'-Tetramethylbenzidine by H2O2, an enzymatic hydrolysis product of cholesterol. And the color transition was more sensitive in the microparticles than in solutions, indicating the microparticles are more applicable for visualized determination. Furthermore, Janus microparticles were employed for multitarget sensing in the two hemespheres, and glucose and cholesterol were detected within the same microparticles without obvious interference. Besides, the particles could be manipulated by an external magnetic field. The glucose and cholesterol levels were measured in human serum utilizing the microparticles, which confirmed the potential application of the microparticles in real sample detection.