4.3 Article

Sample preparation module for bacterial lysis and isolation of DNA from human urine

期刊

BIOMEDICAL MICRODEVICES
卷 11, 期 3, 页码 671-678

出版社

SPRINGER
DOI: 10.1007/s10544-008-9277-1

关键词

Urine; Gram-negative; Bacteria; Lysis; PCR; Urinary tract infection; Microfluidic; Thermoplastic; Cyclic polyolefins

资金

  1. Wallace H. Coulter Foundation

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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 10(1)-10(5) 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 10(1) 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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