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Updated: Aug 1, 2026
Microfluidic Chip Fabrication and Method to Detect Influenza
Published on: March 26, 2013
An integrated nucleic acid detection method based on a microfluidic chip for collection and culture of rice false
Ning Yang1, Yuanyuan Ji1, Aiying Wang2
1School of Electrical and Information Engineering, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
Abstract:
Rice false smut spores (RFSS), which are airborne spores caused by Ustilaginoidea virens (U. virens), not only cause severe yield loss and grain quality reduction, but also produce toxins that are harmful to humans and animals. Nucleic acid detection has become the main method for RFSS monitoring due to its high specificity and sensitivity. However, nucleic acid detection requires multiple steps of spore collection, DNA extraction, nucleic acid amplification and detection, which has a high demand for personnel and is hard to link with other intelligent equipment to achieve automation. Microfluidic chip has become an important approach for integrated detection of pathogens owning to miniaturization and integration in recent years. Yet there is a lack of portable methods that integrate the collection of airborne fungal spores and nucleic acid detection. Because RFSS have thick cell walls and require liquid nitrogen grinding to extract DNA, breaking the walls on-chip is difficult. Therefore, the realization of RFSS wall breaking on-chip is a major difficulty and also a very meaningful study. This study uses RFSS as the research object and provides a novel method of culturing RFSS on-chip to solve the problem of hard wall breaking, realizing the integrated detection of RFSS. The mycelium grown by RFSS germination could be easily broken to release DNA for on-chip detection, which eliminates the need for manual DNA extraction and resolves the issue of difficult wall breaking. This chip can collect RFSS based on the aerodynamic theory and achieve gas-liquid coupling through a simple microvalve structure. A micromixer is constructed to mix the liquid, and then accomplish detection quickly by recombinase polymerase amplification and lateral flow dipsticks (RPA-LFD). The detection sensitivity of this method is 1 × 102-1 × 105 CFU ml-1. It can realize the "sample in and answer out" detection of RFSS due to its simple operation, independence from precision instruments, high sensitivity and specificity. The result shows that it can be used for the early detection of RFSS, has great application prospects and is expected to promote the development of on-site instant detection equipment.
Insights
This study presents a novel microfluidic chip for detecting rice false smut spores (RFSS). The chip integrates spore collection and nucleic acid detection, overcoming challenges in cell wall lysis for automated, on-site monitoring.
Area of Science:
- Agricultural Science
- Biotechnology
- Environmental Science
Background:
- Rice false smut spores (RFSS), caused by *Ustilaginoidea virens*, lead to significant crop losses and produce harmful toxins.
- Current nucleic acid detection methods for RFSS are sensitive but labor-intensive and difficult to automate.
- Existing microfluidic approaches lack integrated airborne fungal spore collection and on-chip cell lysis capabilities.
Purpose of the Study:
- To develop a portable microfluidic chip for integrated collection and detection of airborne RFSS.
- To overcome the challenge of lysing thick-walled RFSS on-chip.
- To enable rapid, automated, and on-site detection of RFSS.
Main Methods:
- Developed a microfluidic chip utilizing aerodynamic principles for RFSS collection.
- Implemented on-chip cultivation of RFSS to facilitate mycelial growth and subsequent cell lysis.
- Integrated gas-liquid coupling, micromixing, recombinase polymerase amplification (RPA), and lateral flow dipsticks (LFD) for detection.
Main Results:
- Successfully cultured RFSS on-chip, enabling easier DNA release and eliminating manual extraction.
- Achieved a detection sensitivity of 1 × 10^2–1 × 10^5 CFU ml^-1 for RFSS.
- Demonstrated a "sample in and answer out" detection system with high specificity and sensitivity.
Conclusions:
- The developed microfluidic chip offers a novel solution for integrated RFSS detection.
- This method simplifies operation, removes the need for precision instruments, and facilitates early detection.
- The technology holds significant promise for developing on-site, instant detection equipment for agricultural pathogens.
