Cell lysis and DNA extraction of gram-positive and gram-negative bacteria from whole blood in a disposable microfluidic chip

• DOI: 10.1039/b905065p
• Publication Date: 2009-06-29T10:30:12Z
• Authors: Madhumita Mahalan...; Hussam Al-Muayad; M. Dominika Kulinski; Dave Altman; Catherine M. Klap...
• Keywords: Gram-Positive Bacteria; Enterococcus faecalis; Gram-Negative Bacteria; Gram

Sepsis caused by gram positive and negative bacteria is the leading cause of death in noncoronary ICUs tenth United States. We have developed a microfluidic sample preparation platform for rapid on-chip detection infectious organisms point-of-care diagnostics. The chips are made robust thermoplastic can be easily multiplexed high throughput applications. Bacteria lysed via hybrid chemical/mechanical method. Once lysed, bacterial DNA isolated using microscale silica bead/polymer composite solid-phase-extraction (SPE) column. Lysis was confirmed off-chip real time PCR. detected both gram-negative (Escherichia coli) gram-positive (Bacillussubtilis Enterococcus faecalis) genomic from microliter scale spiked whole human blood samples. system performs better than it does bacteria, with limits at 102CFU/ml 103–104CFU/ml, respectively. Total extraction times less one hour further decreased altering channel geometry pumping configuration.