Minqiang Bu

A temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a disposable polymer microfluidic device

We present a temperature control method capable of effectively shortening the thermal cycling time of polymerase chain reaction (PCR) in a disposable polymer microfluidic device with an external heater and a temperature sensor. The method employs optimized temperature overshooting and undershooting steps to achieve a rapid ramping between the temperature steps for DNA denaturation, annealing and extension. The temperature dynamics within the microfluidic PCR chamber was characterized and the overshooting and undershooting parameters were optimized using the temperature-dependent fluorescence signal from Rhodamine B. The method was validated with the PCR amplification of mecA gene (162 bp) from methicillin-resistant Staphylococcus aureus bacterium (MRSA), where the time for 30 cycles was reduced from 50 min (without over- and undershooting) to 20 min. (Some figures may appear in colour only in the online journal)

A microfluidic control system with re-usable micropump/valve actuator and injection moulded disposable polymer lab-on-a-slide

A microfluidic control system consisting of micropump/valves with a re-usable pneumatic actuator and a disposable polymer lab-on-a-slide is presented. The lab-on-a-slide was fabricated using low cost methods, such as injection moulding of TOPAS® cyclic olefin copolymer (COC) slide, lamination of different layers of polymer, and ultrasonic welding of TOPAS® lid to the slide. The re-usable pneumatic actuator not only simplifies the design of the lab-on-a-slide and reduces the fabrication cost, but also reduces the possibility of cross contamination during replacement of the disposable lab-on-a-slide. The experiment results show the micropump is capable of providing a wide range of flow rate (20–300 µL/min), and microvalves provides a reliable fluidic control in the different parts of the system.

Rapid sample preparation for detection and identification of avian influenza virus from chicken faecal samples using magnetic bead microsystem

Avian influenza virus (AIV) is an infectious agent of birds and mammals. AIV is causing huge economic loss and can be a threat to human health. Reverse transcriptase polymerase chain reaction (RT-PCR) has been used as a method for the detection and identification of AIV virus. Although RT-PCR is a sensitive method for detection of AIV, it requires sample preparation including separation and purification of AIV and concentrate viral RNA. It is laborious and complex process especially for diagnosis using faecal sample. In this study, magnetic beads were used for immunoseparation of AIV in chicken faecal sample by a magnetic microsystem. Using this system, all the 16 hemagglutinin (H) and 9 neuraminidase (N) subtypes of AIV were separated and detected in spiked faecal samples using RT-PCR, without an RNA extraction step. This rapid sample preparation method can be integrated with a total analysis microsystem and used for diagnosis of AIV.

DNA microarrays immobilized on unmodified plastics in a microfluidic biochip for rapid typing of Avian Influenza Virus

Polymers are widely used for microfluidic systems, but fabrication of microarrays on such materials often requires complicated chemical surface modifications, which hinders the integration of microarrays into microfluidic systems. In this paper, we demonstrate that UV irradiation can be used to directly immobilize poly(T)poly(C)-tagged DNA oligonucleotide probes on non-modified plastic surfaces. This one-step, cost-effective process provides very high immobilization and hybridization efficiencies and is applicable to many different types of polymers. Using this microarray fabrication technique, a portable cyclic olefin copolymer (COC) microarray device containing eight individually addressable microfluidic channels was developed for fast identification of Avian Influenza Virus (AIV) by DNA hybridization. This plastic biochip offers benefits of low fabrication cost and parallel processing of multiple samples, and could be used as a point-of-care device for clinical diagnostics and gene expression analysis.