On-chip genotoxic bioassay based on bioluminescence reporter system using three-dimensional microfluidic network
On-chip genotoxic bioassay based on bioluminescence reporter system using three-dimensional microfluidic network
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DOI：
10.1016/j.aca.2005.12.034
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发表时间：
2006-02
影响因子：
6.2
通讯作者：
Koji Maehana;H. Tani;T. Kamidate
中科院分区：
文献类型：
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作者：
Koji Maehana;H. Tani;T. Kamidate
已经使用感测大肠杆菌菌株进行了基于微芯片的基因毒性生物测定。在该方法中，检测在由硅穿孔微孔阵列芯片和两个聚（二甲基硅氧烷）（PDMS）多微通道芯片构建的三维微流控网络中进行。将具有在作为SOS启动子的umuD转录控制下的萤火虫荧光素酶报告基因的传感菌株放入其中一个PDMS芯片上的通道中并固定在硅微孔中。含有基因毒性物质和荧光素酶底物的样品进入另一个 PDMS 芯片上的通道。使用丝裂霉素 C (MMC) 作为基因毒性物质，研究了芯片上测定的最佳条件。结果，在芯片上立即获得了生物发光强度的剂量依赖性。此外，通过改进芯片上测定方法和条件，成功提高了诱变菌株和非诱导菌株之间的生物发光响应比。对几种众所周知的基因毒性物质进行了芯片检测，并以与传统方法相当的检出限进行了检测，同时缩短了时间。
Microchip-based genotoxic bioassay using sensing Escherichia coli strains has been performed. In this method, the assay was conducted in three-dimensional microfluidic network constructed by a silicon perforated microwell array chip and two poly(dimethylsiloxane) (PDMS) multi-microchannel chips. The sensing strains having firefly luciferase reporter gene under transcriptional control of umuD as an SOS promoter were put into the channels on one of the PDMS chips and immobilized in the silicon microwells. Samples containing genotoxic substances and substrates for luciferase were into the channels on the other PDMS chip. The optimum conditions of the assay in the on-chip format have been investigated using mitomycin C (MMC) as a genotoxic substance. As a result, the dose-dependence of bioluminescence intensity was obtained at once on the chip. Additionally, the response ratios of the bioluminescence between mutagen- and non-induced strains were successfully enhanced by improving the on-chip assay methods and conditions. Several well-known genotoxic substances were subjected to the on-chip assay, and were detected with the detection limits comparable to those in the conventional method with reduced time.