Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip

Hong Chen, Assem Abolmaaty, Peng Li, Constantina Anagnostopoulos, Stefan Dübel, Mohammad Faghri

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

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.

Original languageEnglish (US)
Title of host publicationProceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009
PublisherAmerican Society of Mechanical Engineers (ASME)
Pages733-738
Number of pages6
EditionPART B
ISBN (Print)9780791843857
DOIs
StatePublished - Jan 1 2010
EventASME 2009 International Mechanical Engineering Congress and Exposition, IMECE2009 - Lake Buena Vista, FL, United States
Duration: Nov 13 2009Nov 19 2009

Publication series

NameASME International Mechanical Engineering Congress and Exposition, Proceedings
NumberPART B
Volume12

Other

OtherASME 2009 International Mechanical Engineering Congress and Exposition, IMECE2009
CountryUnited States
CityLake Buena Vista, FL
Period11/13/0911/19/09

Fingerprint

Microfluidics
Escherichia coli
Genes
Phosphatases
Pathogens
Assays
Microscopes
Gels
Biotin
Hot Temperature

All Science Journal Classification (ASJC) codes

  • Mechanical Engineering

Cite this

Chen, H., Abolmaaty, A., Li, P., Anagnostopoulos, C., Dübel, S., & Faghri, M. (2010). Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip. In Proceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009 (PART B ed., pp. 733-738). (ASME International Mechanical Engineering Congress and Exposition, Proceedings; Vol. 12, No. PART B). American Society of Mechanical Engineers (ASME). https://doi.org/10.1115/IMECE2009-11796
Chen, Hong ; Abolmaaty, Assem ; Li, Peng ; Anagnostopoulos, Constantina ; Dübel, Stefan ; Faghri, Mohammad. / Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip. Proceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009. PART B. ed. American Society of Mechanical Engineers (ASME), 2010. pp. 733-738 (ASME International Mechanical Engineering Congress and Exposition, Proceedings; PART B).
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abstract = "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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Chen, H, Abolmaaty, A, Li, P, Anagnostopoulos, C, Dübel, S & Faghri, M 2010, Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip. in Proceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009. PART B edn, ASME International Mechanical Engineering Congress and Exposition, Proceedings, no. PART B, vol. 12, American Society of Mechanical Engineers (ASME), pp. 733-738, ASME 2009 International Mechanical Engineering Congress and Exposition, IMECE2009, Lake Buena Vista, FL, United States, 11/13/09. https://doi.org/10.1115/IMECE2009-11796

Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip. / Chen, Hong; Abolmaaty, Assem; Li, Peng; Anagnostopoulos, Constantina; Dübel, Stefan; Faghri, Mohammad.

Proceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009. PART B. ed. American Society of Mechanical Engineers (ASME), 2010. p. 733-738 (ASME International Mechanical Engineering Congress and Exposition, Proceedings; Vol. 12, No. PART B).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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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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Chen H, Abolmaaty A, Li P, Anagnostopoulos C, Dübel S, Faghri M. Heterogeneous detection of PCR-amplified intimin gene from E. coliO157:H7 via PDMS microfluidic chip. In Proceedings of the ASME International Mechanical Engineering Congress and Exposition 2009, IMECE 2009. PART B ed. American Society of Mechanical Engineers (ASME). 2010. p. 733-738. (ASME International Mechanical Engineering Congress and Exposition, Proceedings; PART B). https://doi.org/10.1115/IMECE2009-11796