Abstract
Silica impregnated polymer monolithic columns may provide a simple method for lysing and extracting DNA from bacteria inside of microfluidic chips. Here we use Escherichia coli as a test organism for a point of care thermoplastic microfluidic module designed to take in a urine sample, mix it with lysis buffer, and perform a hybrid chemical/mechanical lysis and solid phase extraction of nucleic acids from the sample. To demonstrate proof-of-concept, we doped human hematuric urine samples with E. coli at concentrations ranging from 101–105 colony-forming units/mL (CFU/mL) to simulate patient samples. We then performed on-chip lysis and DNA extraction. The bacterial DNA was amplified using real-time PCR demonstrating lysis and isolation down to 101 CFU/mL. Results were comparable to a commercial kit at higher concentrations and performed better at recovering DNA at lower concentrations.
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We thank the Wallace H. Coulter Foundation for funding this research and H. Muayad for help with image analysis.
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Authors M. Dominika Kulinski and Madhumita Mahalanabis contributed equally to this manuscript.
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Kulinski, M.D., Mahalanabis, M., Gillers, S. et al. Sample preparation module for bacterial lysis and isolation of DNA from human urine. Biomed Microdevices 11, 671–678 (2009). https://doi.org/10.1007/s10544-008-9277-1
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DOI: https://doi.org/10.1007/s10544-008-9277-1