Abstract
BackgroundMicrofluidic chip detection technology is considered a potent tool for many bioanalytic applications. Rapid detection of foodborne pathogens in the early stages is imperative to prevent the outbreak of foodborne diseases, known as a severe threat to human health. Conventional bacterial culture methods for detecting foodborne pathogens are time-consuming, laborious, and lacking in pathogen diagnosis. To overcome this problem, we have created an embedded paper-based microchip based on isothermal loop amplification (LAMP), which can rapidly and sensitively detect foodborne pathogens.ResultsWe embed paper impregnated with LAMP reagent and specific primers in multiple reaction chambers of the microchip. The solution containing the target pathogen was injected into the center chamber and uniformly distributed into the reaction chamber by centrifugal force. The purified DNA of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus, and Vibrio parahaemolyticus has been successfully amplified and directly detected on the microchip. The E. coli O157:H7 DNA was identified as low as 0.0134 ng μL− 1. Besides, the potential of this microchip in point-of-care testing was further tested by combining the on-chip sample purification module and using milk spiked with Salmonella spp.. The pyrolyzed milk sample was filtered through a polydopamine-coated paper embedded in the inside of the sample chamber. It was transported to the reaction chamber by centrifugal force for LAMP amplification. Then direct chip detection was performed in the reaction chamber embedded with calcein-soaked paper. The detection limit of Salmonella spp. in the sample measured by the microchip was approximately 12 CFU mL− 1.ConclusionThe paper embedded LAMP microchip offers inexpensive, user-friendly, and highly selective pathogen detection capabilities. It is expected to have great potential as a quick, efficient, and cost-effective solution for future foodborne pathogen detection.
Highlights
Microfluidic chip detection technology is considered a potent tool for many bioanalytic applications
Optimizing loop amplification (LAMP) reaction conditions To optimize the conditions of the LAMP reaction, firstly, the reaction temperature was set to 60 °C, 65 °C, and 70 °C, and the reaction time was set to 15 min, 20 min, 25 min, 30 min, and 40 min
We found that the temperature of the ceramic heating plate can be maintained at 65 °C ± 0.1 °C for 30 min, which is enough to complete a LAMP reaction process and meet the temperature requirements of the LAMP reaction on the microchip
Summary
Microfluidic chip detection technology is considered a potent tool for many bioanalytic applications. Conventional bacterial culture methods for detecting foodborne pathogens are time-consuming, laborious, and lacking in pathogen diagnosis. Conventional microbial detection methods depend upon culturing organisms in selective media followed by microbial identification employed morphological, biochemical, or immunological characteristics [5]. These methods typically require 3 to 5 days to confirm pathogenic microorganisms [6]. This time frame does not meet the need for rapid clinical diagnosis or identification in field food security investigations, especially in cases of acute infection. It is essential to develop a rapid and sensitive method that can be broadly applied for detecting foodborne pathogenic microorganisms
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