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A directional self-priming continuous-driven compartmentalized microfluidic chip for multiplexed pathogen detection
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DOI：10.1039/D5AN00640F.png)
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Abstract
En 中文
Rapid and efficient screening of foodborne pathogens is crucial for preventing bacterial spread and food poisoning. However; developing a multi-detection method that is easy to operate; offers good stability; and achieves high efficiency remains an enormous challenge. Existing multiplexed nucleic acid detection methods suffer from complex designs; leading to complicated operations; and non-robust sample introduction; causing primer/probe crosstalk and false-positive results. To address these issues; we developed a directional self-priming continuous-driven multiplexed detection microfluidic chip based on a snowflake shaped-inspired design with six branches; and each branch carried a unique functional input inlet for different specific targets; enabling simultaneous detection of multiple targets from one sample using monochrome fluorescence. Based on the PDMS negative pressure storage capacity; this chip explored the self-priming method that enables the continuous loading of primers/probes and samples without requiring an external power source; effectively addressing key challenges in multiplexed detection. This chip with specially designed outlet-free and symmetric inverted T-shaped microchannels allows for multi-step reagent loading and 100% compartmentalization without reagent loss; significantly reducing reagent consumption and sample loss compared to current methods. For further validation; we tested three types of foodborne pathogens (Salmonella enterica; Staphylococcus aureus; and Listeria monocytogenes) to demonstrate the chip's multiplexed detection capability; achieving a notably high sensitivity of detection as low as 10 copies per μL. Our snowflake-shaped microfluidic chip platform shows great potential for future applications in point-of-care (POC) settings; offering a robust and efficient solution for detecting foodborne pathogens.
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