Sample preparation module for bacterial lysis and isolation of DNA from human urine.
Sample preparation module for bacterial lysis and isolation of DNA from human urine.
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DOI：
10.1007/s10544-008-9277-1
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发表时间：
2009-06
影响因子：
2.8
通讯作者：
Klapperich, Catherine M.
中科院分区：
文献类型：
--
作者：
Kulinski, M. Dominika;Mahalanabis, Madhumita;Gillers, Sara;Zhang, Jane Y.;Singh, Satish;Klapperich, Catherine M.
关键词：
硅浸渍聚合物整体柱可为微流控芯片内细菌的DNA裂解和提取提供一种简单的方法。在这里，我们使用大肠杆菌作为护理点热塑性微流控模块的测试生物，设计用于提取尿液样本，将其与裂解缓冲液混合，并从样本中进行混合化学/机械裂解和固相萃取核酸。为了证明概念验证，我们在人类血尿样本中掺入浓度为101-105集落形成单位/mL （CFU/mL）的大肠杆菌来模拟患者样本。然后我们进行片上裂解和DNA提取。利用实时荧光定量PCR扩增细菌DNA，证实裂解和分离的浓度为101 CFU/mL。结果在较高浓度下与商业试剂盒相当，在较低浓度下恢复DNA时表现更好。
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 concen trations and performed better at recovering DNA at lower concentrations.
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影响因子：
7.4
作者：
Bhattacharyya, A;Klapperich, CM
通讯作者：
Klapperich, CM
影响因子：
2.9
作者：
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作者：
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通讯作者：
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