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An integrated microfluidic system using mannose-binding lectin for bacteria isolation and biofilm-related gene detection
Authors:Ju-Ching Yu  Chih-Chien Hu  Wen-Hsin Chang  Pei-Chun Chen  Mel S Lee  Kuo-Ti Peng  Gwo-Bin Lee
Affiliation:1.Department of Power Mechanical Engineering,National Tsing Hua University,Hsinchu,Taiwan;2.Department of Orthopedics, Chang Gung Memorial Hospital, Linkou,Chang Gung University,Taoyüan,Taiwan;3.Department of Orthopaedic Surgery,Chiayi Chang Gung Memorial Hospital,Chiayi,Taiwan;4.Department of Orthopaedic Surgery,Kaohsiung Chang Gung Memorial Hospital,Kaohsiung,Taiwan;5.Institute of Biomedical Engineering,National Tsing Hua University,Hsinchu,Taiwan;6.Institute of NanoEngineering and Microsystems,National Tsing Hua University,Hsinchu,Taiwan
Abstract:Molecular diagnosis of biofilm-related genes (BRGs) in common bacteria that cause periprosthetic joint infections may provide crucial information for clinicians. In this study, several BRGs, including ica, fnbA, and fnbB, were rapidly detected (within 1 h) with a new integrated microfluidic system. Mannose-binding lectin (MBL)-coated magnetic beads were used to isolate these bacteria, and on-chip nucleic acid amplification (polymerase chain reaction, PCR) was then performed to detect BRGs. Both eukaryotic and prokaryotic MBLs were able to isolate common bacterial strains, regardless of their antibiotic resistance, and limits of detection were as low as 3 and 9 CFU for methicillin-resistant Staphylococcus aureus and Escherichia coli, respectively, when using a universal 16S rRNA PCR assay for bacterial identification. It is worth noting that the entire process including bacteria isolation by using MBL-coated beads for sample pre-treatment, on-chip PCR, and fluorescent signal detection could be completed on an integrated microfluidic system within 1 h. This is the first time that an integrated microfluidic system capable of detecting BRGs by using MBL as a universal capturing probe was reported. This integrated microfluidic system might therefore prove useful for monitoring profiles of BRGs and give clinicians more clues for their clinical judgments in the near future.
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