Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip
Date of Original Version
E. coli O157:H7 strains represent the most important group of food-borne pathogens. PCR-amplified intimin gene of pathogenic E. coli O 157:H7 was detected heterogeneously via a microfluidic chip that consists of streptavidin-coated nanoliter chambers. Biotinylated primers and digoxigenin labeled deoxyuridine triphosphate (dUTP) were incorporated into the amplified intimin (eaeA) gene by an off-chip PCR thermal cycler, The amplified products were injected into the chip where they were immobilized via streptavidin-biotin interaction. Detection of the products using alkaline phosphatase (AP) conjugated anti-digoxigenin was performed with an epi-fluorescent microscope. This assay was capable of detecting 0.06 ng/μL biotin-digoxigenin-dsDNA conjugate distinctly, which is a hundred fold more sensitive than the traditional detection by agarose gel. Copyright © 2010 by ASME.
ASME International Mechanical Engineering Congress and Exposition, Proceedings
Chen, Hong, Assem Abolmaaty, Peng Li, Constantina Anagnostopoulos, Stefan Dübel, and Mohammad Faghri. "Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip." ASME International Mechanical Engineering Congress and Exposition, Proceedings 12, PART B (2010): 733-738. doi:10.1115/IMECE2009-11796.