A new microfluidic reaction chip capable of mixing, transporting and controlling reactions has been developed for the size-tunable synthesis of gold nanoparticles. This chip allows for an accelerated and efficient approach for the synthesis of gold nanoparticles. The microfluidic reaction chip is made by computer-numerically controlled machining and PDMS casting processes, which integrate a micro-mixer, a normally closed valve and a micro-pump onto a single chip. The micro-mixer is capable of generating a vortex-type flow field, which achieves a mixing efficiency as high as 95% within 1 s. Successful synthesis of dispersed gold nanoparticles has been demonstrated within an 83% shorter period of time (13 min), as compared to traditional methods (around 2 h). By using different volumes of reagents, the dispersed gold nanoparticles are found to have average diameters of 19, 28, 37 and 58 nm. The optical absorption spectra indicate that these synthesized nanoparticles have different surface plasmon resonance peaks, which are 521, 525, 530 and 537 nm, respectively. The development of this microfluidic reaction system holds promise for the synthesis of functional nanoparticles for further biomedical applications.
Arthroplasty is a general approach for improving the life quality for patients with degenerative or injured joints. However, post-surgery complications including periprosthetic joint infection (PJI) poses a serious drawback to the procedure. Several methods are available for diagnosing PJI, but they are time-consuming or have poor sensitivity and specificity. Alternatively, reverse-transcription PCR can detect live bacteria and reduce false-positive results but cannot avoid the cumbersome RNA handling and human contamination issues. In response, an integrated microfluidic system capable of detecting live bacteria from clinical PJI samples within 55 minutes is developed in this study. This system employs an ethidium monoazide (EMA)-based assay and a PCR with universal bacterial primers and probes to isolate and detect only the live bacteria that commonly cause PJI. The experimental results indicated that the developed system can detect bacteria in human joint fluids with a detection limit of 10(4) colony formation unit mL(-1). Furthermore, nine clinical samples were analyzed using the microfluidic system. The results obtained from the microfluidic system were negative for all culture-negative cases, indicating that the proposed system can indeed reduce false-positive results. In addition, experimental results showed that the EMA sample pre-treatment process was crucial for successful detection of live bacteria. The culture-positive cases were diagnosed as positive by the proposed system only when the clinical samples were treated with EMA immediately after being sampled from patients. Based on these promising results, the developed microfluidic system can be a useful tool to detect PJI and potentially be applied in other clinical situations.
Vancomycin-resistant (VRE) is a kind of enterococci, which shows resistance toward antibiotics. It may last for a long period of time and meanwhile transmit the vancomycin-resistant gene () to other bacteria. In the United States alone, the resistant rate of to vancomycin increased from a mere 0.3% to a whopping 40% in the past two decades. Therefore, timely diagnosis and control of VRE is of great need so that clinicians can prevent patients from becoming infected. Nowadays, VRE is diagnosed by antibiotic susceptibility test or molecular diagnosis assays such as matrix-assisted laser desorption ionization/time-of-flight mass spectrometry and polymerase chain reaction. However, the existing diagnostic methods have some drawbacks, for example, time-consumption, no geneticinformation, or high false-positive rate. This study reports an integrated microfluidic system, which can automatically identify the vancomycin resistant gene () from live bacteria in clinical samples. A new approach using ethidium monoazide, nucleic acid specific probes, low temperature chemical lysis, and loop-mediated isothermal amplification (LAMP) has been presented. The experimental results showed that the developed system can detect the gene from live in joint fluid samples with detection limit as low as 10 colony formation units/reaction within 1 h. This is the first time that an integrated microfluidic system has been demonstrated to detect gene from live bacteria by using the LAMP approach. With its high sensitivity and accuracy, the proposed system may be useful to monitor antibiotic resistance genes from live bacteria in clinical samples in the near future.
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