Total integrated slidable and valveless solid phase extraction-polymerase chain reaction-capillary electrophoresis microdevice for mini Y chromosome short tandem repeat genotyping

Biosens Bioelectron. 2016 Apr 15:78:489-496. doi: 10.1016/j.bios.2015.11.079. Epub 2015 Dec 2.

Abstract

A fully integrated slidable and valveless microsystem, which performs solid phase DNA extraction (SPE), micro-polymerase chain reaction (μPCR) and micro-capillary electrophoresis (μCE) coupled with a portable genetic analyser, has been developed for forensic genotyping. The use of a slidable chip, in which a 1 μL-volume of the PCR chamber was patterned at the center, does not necessitate any microvalves and tubing systems for fluidic control. The functional micro-units of SPE, μPCR, and μCE were fabricated on a single glass wafer by conventional photolithography, and the integrated microdevice consists of three layers: from top to bottom, a slidable chip, a channel wafer in which a SPE chamber, a mixing microchannel, and a CE microchannel were fabricated, and a Ti/Pt resistance temperature detector (RTD) wafer. The channel glass wafer and the RTD glass wafer were thermally bonded, and the slidable chip was placed on the designated functional unit. The entire process from the DNA extraction using whole human blood sample to identification of target Y chromosomal short tandem repeat (STR) loci was serially carried out with simply sliding the slidable chamber from one to another functional unit. Monoplex and multiplex detection of amelogenin and mini Y STR loci were successfully analysed on the integrated slidable SPE-μPCR-μCE microdevice by using 1 μL whole human blood within 60 min. The proposed advanced genetic analysis microsystem is capable of point-of-care DNA testing with sample-in-answer-out capability, more importantly, without use of complicated microvalves and microtubing systems for liquid transfer.

Keywords: Capillary electrophoresis; Integrated microdevice; Mini Y STR typing; Polymerase chain reaction; Solid phase DNA extraction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques*
  • Chromosomes, Human, Y / chemistry
  • Chromosomes, Human, Y / genetics
  • DNA / blood*
  • DNA / chemistry
  • Electrophoresis, Capillary*
  • Genotype
  • Humans
  • Lab-On-A-Chip Devices
  • Microsatellite Repeats / genetics*
  • Point-of-Care Systems
  • Polymerase Chain Reaction
  • Solid Phase Extraction

Substances

  • DNA