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


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 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.

SUBMITTER: Kulinski MD 

PROVIDER: S-EPMC4442481 | biostudies-literature | 2009 Jun

REPOSITORIES: biostudies-literature

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

Kulinski M Dominika MD   Mahalanabis Madhumita M   Gillers Sara S   Zhang Jane Y JY   Singh Satish S   Klapperich Catherine M CM  

Biomedical microdevices 20090601 3


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.  ...[more]

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