Dielectrophoresis-based Microfluidic Biochip for Rapid Diagnosis of Dengue Infection

  • 安 迪

Student thesis: Doctoral Thesis

Abstract

The infectious diseases have caused a threat and burden for the humankind and the effort for the discovery and development of a diagnostic tool is continuously required along with the prevalence of diseases Dengue is an emerging viral infection increasing annually worldwide A rapid sensitive and accurate result of the dengue infection diagnosis are still a crucial issue to provide a proper treatment and management In this report the proof of concept of utilizing a method based on microfluidic dielectrophoresis (DEP) chip for diagnosis of dengue virus (DENV) infection is introduced The sensing principle is that hydrodynamic and DEP forces allow the immuno-reaction on the designed chip The subject of this thesis is divided into two topics related to the target of detection that is the dengue virus particle and recombinant dengue virus non-structural protein 1 (rNS1) I Detection of dengue virus particle in vitro The DEP force was employed to capture the modified beads (mouse anti-flavivirus monoclonal antibody-coated beads) flowing in the microfluidic chip and the DENV modified with fluorescence label as the detection target can be then captured on the modified beads by immunoreaction The fluorescent signal obtained was then quantified by image processing software The platform can accelerate an immuno-reaction time in which the on-chip detection time was 5 min and demonstrating an ability for DENV detection as low as 104 PFU/mL II Detection of recombinant dengue virus non-structural protein 1 (rNS1) In this part the beads was immobilized by anti-NS1 specific antibodies (DN5C6 monoclonal antibody-immobilized beads) and the rNS1 was labeled by fluorescent probe via biotin-streptavidin binding as the target detection By immunoreaction mechanism allowed on the chip the platform enables to quantify rNS1 based on fluorescence signal with a detection limit of 10 ng/mL within 10 min of on-chip detection time Furthermore the required volume of DENV and rNS1 samples used were reduced become ~15 ?L and the chip was reusable (>50x) Overall this platform provides a rapid detection (5-10 min) with a low sample volume compared to conventional methods In summary this proof of concept with regard to a microfluidic dielectrophoresis chip thus shows the potential of immunofluorescence based-assay applications to meet diagnostic needs
Date of Award2017 Sep 13
Original languageEnglish
SupervisorHsien-Chang Chang (Supervisor)

Cite this

'