Genotyping of single nucleotide polymorphisms by melting curve analysis using thin film semi-transparent heaters integrated in a lab-on-foil system

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Anna Ohlander - , Fraunhofer Research Institution for Microsystems and Solid State Technologies (EMFT) (Autor:in)
  • Caterina Zilio - , National Research Council of Italy (Autor:in)
  • Tobias Hammerle - , Fraunhofer Research Institution for Microsystems and Solid State Technologies (EMFT) (Autor:in)
  • Sergey Zelenin - , KTH Royal Institute of Technology (Autor:in)
  • Gerhard Klink - , Fraunhofer Research Institution for Microsystems and Solid State Technologies (EMFT) (Autor:in)
  • Marcella Chiari - , National Research Council of Italy (Autor:in)
  • Karlheinz Bock - , Professur für Aufbau- und Verbindungstechnik der Elektronik, Fraunhofer Research Institution for Microsystems and Solid State Technologies (EMFT), Technische Universität Berlin (Autor:in)
  • Aman Russom - , KTH Royal Institute of Technology (Autor:in)

Abstract

The recent technological advances in micro/nanotechnology present new opportunities to combine microfluidics with microarray technology for the development of small, sensitive, single-use, point-of-care molecular diagnostic devices. As such, the integration of microarray and plastic microfluidic systems is an attractive low-cost alternative to glass based microarray systems. This paper presents the integration of a DNA microarray and an all-polymer microfluidic foil system with integrated thin film heaters, which demonstrate DNA analysis based on melting curve analysis (MCA). A novel micro-heater concept using semi-transparent copper heaters manufactured by roll-to-roll and lift-off on polyethylene naphthalate (PEN) foil has been developed. Using a mesh structure, heater surfaces have been realized in only one single metallization step, providing more efficient and homogenous heating characteristics than conventional meander heaters. A robust DNA microarray spotting protocol was adapted on Parylene C coated heater-foils, using co-polymer poly(DMA-NAS-MAPS) to enable covalent immobilization of DNA. The heaters were integrated in a microfluidic channel using lamination foils and MCA of the spotted DNA duplexes showed single based discrimination of mismatched over matched target DNA-probes. Finally, as a proof of principle, we perform MCA on PCR products to detect the Leu7Pro polymorphism of the neutropeptide Y related to increased risk of Type II diabetes, BMI and depression.

Details

OriginalspracheEnglisch
Seiten (von - bis)2075-2082
Seitenumfang8
FachzeitschriftLab on a chip
Jahrgang13
Ausgabenummer11
PublikationsstatusVeröffentlicht - 7 Juni 2013
Peer-Review-StatusJa

Externe IDs

ORCID /0000-0002-0757-3325/work/139064943