Rapid detection of pathogens using antibody-coated microbeads with bioluminescence in microfluidic chips |
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Authors: | Xiao Guan Hui-jing Zhang Yin-nan Bi Li Zhang Dun-ling Hao |
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Institution: | (1) Department of Analytical Chemistry, Faculty of Laboratory Medicine, Third Military Medical University, Chongqing, China, 400038; |
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Abstract: | Detection of pathogens was demonstrated in a polydimethylsiloxane (PDMS)/glass microfluidic chip with which microbead-based
immunoseparation platform and the bioluminescence technology were integrated. Escherichia coli (E. coli) O157:H7 was used as the model bacteria. The microchamber in microfluidic chip was filled with glass beads coated with antibodies
which could capture specific organism, and the capture efficiency of the chip for the bacteria was about 91.75%∼95.62%. Then
the concentration of bacteria was determined by detecting adenosine triphosphate (ATP) employing bioluminescence reaction
of firefly luciferin-lucifera-ATP on chip. The method allowed reliable detection of E. coli O157:H7 concentrations from 3.2 × 101 cfu/μL to 3.2 × 105 cfu/μL within 20 min. This research demonstrated excellent reproducibility, stability, and specificity, and could accurately
detect the pathogenic bacteria in food samples. The microfluidic chip and the equipments used in this method are easy to miniaturize,
thus the method has great potential to be developed to a portable device for rapid detection of pathogens. |
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