One of the main challenges in the diagnosis of infectious diseases is the need for rapid and accurate detection of the causative pathogen in any setting. Rapid diagnosis is key to avoiding the spread of the disease, to allow proper clinical decisions to be made in terms of patient treatment, and to mitigate the rise of drug-resistant pathogens. In the last decade, significant interest has been devoted to the development of point-of-care reverse transcription polymerase chain reaction (PCR) platforms for the detection of RNA-based viral pathogens. We present the development of a microfluidic, real-time, fluorescence-based, continuous-flow reverse transcription PCR system. The system incorporates a disposable microfluidic chip designed to be produced industrially with cost-effective roll-to-roll embossing methods. The chip has a long microfluidic channel that directs the PCR solution through areas heated to different temperatures. The solution first travels through a reverse transcription zone where RNA is converted to complementary DNA, which is later amplified and detected in real time as it travels through the thermal cycling area. As a proof of concept, the system was tested for Ebola virus detection. Two different master mixes were tested, and the limit of detection of the system was determined, as was the maximum speed at which amplification occurred. Our results and the versatility of our system suggest its promise for the detection of other RNA-based viruses such as Zika virus or chikungunya virus, which constitute global health threats worldwide. Graphical abstract Photograph of the RT-PCR thermoplastic chip.

In this paper, we present a portable and low cost point-of-care (POC) PCR system for quantitative detection of pathogens. Our system is based on continuous flow PCR which maintains fixed temperatures zones and pushes the PCR solution between two heated areas allowing for faster heat transfer and as a result, a faster PCR. The PCR system is built around a 46.0 mm × 30.9 mm × 0.4 mm disposable thermoplastic chip. In order to make the single-use chip economically viable, it was manufactured by hot embossing and was designed to be compatible with roll-to-roll embossing for large scale production. The prototype instrumentation surrounding the chip includes two heaters, thermal sensors, and an optical system. The optical system allows for pathogen detection via real time fluorescence measurements. FAM probes were used as fluorescent reporters of the amplicons generated during the PCR. To demonstrate the function of the chip, two infectious bacteria targets were selected: Chlamydia trachomatis and Escherichia coli O157:H7. For both bacteria, the limit of detection of the system was determined, PCR efficiencies were calculated, and different flow velocities were tested. We have demonstrated successful detection for these two bacterial pathogens highlighting the versatility and broad utility of our portable, low-cost, and rapid PCR diagnostic device.

The 5-year survival rate for ovarian cancer is only 45% largely due to lack of effective screening methods. Current methods include palpation, transvaginal ultrasound, and the CA-125 blood test. Finding disease reliably and at an early stage increase survival to 92%. We have designed and built a 0.7 mm endoscope for the early detection of ovarian cancer. Inserted transvaginally through the working channel of a hysteroscope, the falloposcope creates a minimally invasive procedure for the screening of high risk women. To improve the ease-of-use and safety of falloposcope deployment, we are working to create an everting balloon. Currently, the falloposcope would require a skilled user to operate due to the challenging anatomy of the fallopian tubes -a small opening from the uterus (< 1 mm), tortuous path, and delicate lumenal features. A balloon delivery system would gently open the fallopian tube and guide the falloposcope down the center of lumen. We show balloon design and discuss integration with the falloposcope prototype. We test possible mechanical damage to the tissue due to scraping, puncture, or overstretching. Successful introduction of the everting balloon to simplify falloposcope delivery could expand screening beyond specialized centers to smaller clinical locations.