Accurate quantification of DNA using on-site PCR (osPCR) by characterizing DNA amplification at single-molecule resolution.

Despite the need in various applications, accurate quantification of nucleic acids still remains a challenge. The widely-used qPCR has reduced accuracy at ultralow template concentration and is susceptible to nonspecific amplifications. The more recently developed dPCR is costly and cannot handle high-concentration samples. We combine the strengths of qPCR and dPCR by performing PCR in silicon-based microfluidic chips and demonstrate high quantification accuracy in a large concentration range. Importantly, at low template concentration, we observe on-site PCR (osPCR), where only certain sites of the channel show amplification. The sites have almost identical ct values, showing osPCR is a quasi-single molecule phenomenon. Using osPCR, we can measure both the ct values and the absolute concentration of templates in the same reaction. Additionally, osPCR enables identification of each template molecule, allowing removal of nonspecific amplification during quantification and greatly improving quantification accuracy. We develop sectioning algorithm that improves the signal amplitude and demonstrate improved detection of COVID in patient samples.

Ultrafast Nucleic Acid Detection Equipment with Silicon-Based Microfluidic Chip.

Recently, infectious diseases, such as COVID-19, monkeypox, and Ebola, are plaguing human beings. Rapid and accurate diagnosis methods are required to preclude the spread of diseases. In this paper, an ultrafast polymerase chain reaction (PCR) equipment is designed to detect virus. The equipment consists of a silicon-based PCR chip, a thermocycling module, an optical detection module, and a control module. Silicon-based chip, with its thermal and fluid design, is used to improve detection efficiency. A thermoelectric cooler (TEC), together with a computer-controlled proportional-integral-derivative (PID) controller, is applied to accelerate the thermal cycle. A maximum of four samples can be tested simultaneously on the chip. Two kinds of fluorescent molecules can be detected by optical detection module. The equipment can detect viruses with 40 PCR amplification cycles in 5 min. The equipment is portable, easily operated, and low equipment cost, which shows great potential in epidemic prevention.

A Modified Surgical Face Mask to Improve Protection and Wearing Comfort

Wearing face masks is essential for reducing infection during the COVID-19 pandemic. However, ordinary surgical face masks can provide only moderate protection. The N95 face masks should provide sufficient protection but may impose complaints about breathing difficulty or even impair respiratory health. This investigation proposed a novel face mask modified from the surgical face mask to improve both protection and comfort. The filter material of the surgical face mask was covered and sealed on a cardboard support frame but with openings for air permeating through. The modified face masks were worn by a test subject for measuring the air contents inside the face masks. The protection performance was evaluated by the overall PM1 filtration efficiency. The concentrations of CO2, O2, N2, and water vapor were adopted to evaluate the breathing comfort. The performance of the proposed face mask was compared with the market-available surgical and N95 face masks. In addition, CFD modeling was adopted to investigate the dynamic air exchange of the face mask with respiration and the surrounding air. Impacts of the air sampling tube positions on the measurement results were also examined. The results revealed that the overall PM1 filtration efficiency of the modified face mask could reach 96.2%, which was much higher than that of the surgical face mask and only slightly lower than the N95 face mask. As compared with the N95 face mask, the modified mask reduced the respiratory flow resistance and the concentrations of CO2 and water vapor and thus increased the O2 content and breathing comfort.