用于细菌检测的集成微流控芯片——概念验证

An integrated microfluidic chip for the detection of bacteria - A proof of concept.

作者信息

Guo Zhe, Yu Ting, He Jiarui, Liu Fen, Hao Hualong, Zhao Yang, Wen Jiabin, Wang Qi

机构信息

Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China.

Beijing Institute of Respiratory Medicine, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100069, China.

出版信息

Mol Cell Probes. 2015 Aug;29(4):223-7. doi: 10.1016/j.mcp.2015.05.005. Epub 2015 May 13.

DOI:10.1016/j.mcp.2015.05.005

PMID:25979593

Abstract

We designed a microfluidic chip as a proof of concept for the detection of bacterial DNA. The chip was fabricated with poly-dimethylsiloxane (PDMS). It included a solid phase extraction (SPE) chamber, two separate channels and multiple loop-mediated isothermal amplification (LAMP) chambers. Three bacterial strains (Escherichia coli O157:H7, methicillin-resistant Staphylococcus aureus and methicillin-sensitive S. aureus) were used to test the feasibility of the device. LAMP products were examined directly using a UV light and verified by agarose gel electrophoresis. Using this chip, we successfully detected E. coli O157:H7, MSSA and MRSA in less than 2 h. The detection limit for genes rfbE, spa and mecA (specific to E. coli O157:H7, MSSA and MRSA, respectively) was <10(2) CFU/100 μl. Further work is required to refine this approach and rigorously assess its analytical and diagnostic specificity and sensitivity.

摘要

我们设计了一种微流控芯片作为检测细菌DNA概念验证。该芯片由聚二甲基硅氧烷（PDMS）制成。它包括一个固相萃取（SPE）腔室、两条独立通道和多个环介导等温扩增（LAMP）腔室。使用三种细菌菌株（大肠杆菌O157:H7、耐甲氧西林金黄色葡萄球菌和甲氧西林敏感金黄色葡萄球菌）来测试该装置的可行性。LAMP产物直接用紫外光检测并用琼脂糖凝胶电泳验证。使用该芯片，我们在不到2小时内成功检测出大肠杆菌O157:H7、甲氧西林敏感金黄色葡萄球菌和耐甲氧西林金黄色葡萄球菌。rfbE、spa和mecA基因（分别特异性针对大肠杆菌O157:H7、甲氧西林敏感金黄色葡萄球菌和耐甲氧西林金黄色葡萄球菌）的检测限<10(2) CFU/100 μl。需要进一步开展工作来完善这种方法，并严格评估其分析和诊断特异性及灵敏度。