Kulinski M Dominika, Mahalanabis Madhumita, Gillers Sara, Zhang Jane Y, Singh Satish, Klapperich Catherine M
Department of Mechanical Engineering, Boston University, 44 Cummington St., Boston, MA, USA.
Department of Biomedical Engineering, Boston University, 44 Cummington St., Boston, MA, USA.
Biomed Microdevices. 2009 Jun;11(3):671-678. doi: 10.1007/s10544-008-9277-1.
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.
二氧化硅浸渍的聚合物整体柱可能为从微流控芯片内的细菌中裂解和提取DNA提供一种简单方法。在这里，我们使用大肠杆菌作为测试生物体，用于一种即时护理热塑性微流控模块，该模块设计用于采集尿液样本，将其与裂解缓冲液混合，并对样本中的核酸进行化学/机械混合裂解和固相提取。为了证明概念验证，我们在人血尿尿液样本中掺入浓度范围为10(1)-10(5) 菌落形成单位/毫升（CFU/毫升）的大肠杆菌，以模拟患者样本。然后我们进行了芯片上的裂解和DNA提取。使用实时PCR对细菌DNA进行扩增，证明裂解和分离可低至10(1) CFU/毫升。在较高浓度下，结果与商业试剂盒相当，在较低浓度下回收DNA的表现更好。
