采用集成非晶硅氢化探测器的微流控基因分析

Microfluidic genetic analysis with an integrated a-Si:H detector.

作者信息

Kamei T, Toriello N M, Lagally E T, Blazej R G, Scherer J R, Street R A, Mathies R A

机构信息

Department of Chemistry, University of California, Berkeley, CA 94720, USA.

出版信息

Biomed Microdevices. 2005 Jun;7(2):147-52. doi: 10.1007/s10544-005-1595-y.

DOI:10.1007/s10544-005-1595-y

PMID:15940430

Abstract

We have developed an integrated hydrogenated amorphous silicon (a-Si:H) fluorescence detector for microfluidic genetic analysis. It consists of a half-ball lens, a ZnS/YF3 multilayer optical interference filter with a pinhole, and an annular a-Si:H PIN photodiode allowing the laser excitation to pass up through the central aperture in the photodiode and the filter. Microfluidic separations of multiplex PCR products generated from methicillin-resistant/sensitive Staphylococcus aureus (MRSA/MSSA) DNA on microfluidic capillary electrophoresis (CE) devices are successfully detected with the integrated detector. Similarly, multiplex PCR amplicons from the kanamycin resistant and K12 serotype-specific genes of E. coli cells are detected. The direct detection of multiplex PCR amplicons indicates that the fluorescence detector can be successfully coupled with current microfluidic PCR-CE platforms. This work establishes that the integrated a-Si:H detector provides relevant limits of detection for point-of-care genetic and pathogen analysis with microfluidic devices.

摘要

我们开发了一种用于微流控基因分析的集成氢化非晶硅（a-Si:H）荧光探测器。它由一个半球形透镜、一个带有针孔的ZnS/YF3多层光学干涉滤光片以及一个环形a-Si:H PIN光电二极管组成，该光电二极管允许激光激发光通过光电二极管和滤光片中的中心孔径。利用该集成探测器成功检测了在微流控毛细管电泳（CE）装置上对耐甲氧西林/敏感金黄色葡萄球菌（MRSA/MSSA）DNA产生的多重PCR产物进行的微流控分离。同样，也检测到了来自大肠杆菌细胞卡那霉素抗性和K12血清型特异性基因的多重PCR扩增子。多重PCR扩增子的直接检测表明，该荧光探测器可以成功地与当前的微流控PCR-CE平台耦合。这项工作表明，集成的a-Si:H探测器为使用微流控装置进行即时基因和病原体分析提供了相关的检测限。