TY - GEN
T1 - Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip
AU - Chen, Hong
AU - Abolmaaty, Assem
AU - Li, Peng
AU - Anagnostopoulos, Constantina
AU - Dübel, Stefan
AU - Faghri, Mohammad
PY - 2010
Y1 - 2010
N2 - 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.
AB - 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.
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U2 - 10.1115/IMECE2009-11796
DO - 10.1115/IMECE2009-11796
M3 - Conference contribution
AN - SCOPUS:77954248690
SN - 9780791843857
T3 - ASME International Mechanical Engineering Congress and Exposition, Proceedings
SP - 733
EP - 738
BT - Proceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009
PB - American Society of Mechanical Engineers (ASME)
T2 - ASME 2009 International Mechanical Engineering Congress and Exposition, IMECE2009
Y2 - 13 November 2009 through 19 November 2009
ER -