Rapid screening of phenylketonuria using a CD microfluidic device.

Abstract

We herein present a compact disc (CD) microfluidic chip based hybridization assay for phenylketonuria (PKU) screening. This CD chip is composed of a polydimethylsiloxane (PDMS) top layer containing 12 DNA hybridization microchannels, and a glass bottom layer with hydrogel pad conjugated DNA oligonucleotides. Reciprocating flow was generated on the CD chip through a simple rotation-pause operation to facilitate rapid DNA hybridization. When rotated the CD chip, the sample solution was driven into the hybridization channel by centrifugal force. When stopped the CD chip, the sample plug was pulled backward through the channel by capillary force. The hybridization assay was firstly validated with control samples and was then used to analyze 30 clinical samples from pregnant women with suspected PKU fetus. The on-chip DNA hybridization was completed in 15 min with a sample consumption as low as 1.5μL, and the limit-of-detection (LOD) of DNA template was 0.7ng/μL. Among the 30 samples tested, V245V mutation was identified in 4 cases while R243Q mutation was detected in one case. Results of the hybridization assay were confirmed by DNA sequencing. This CD-chip based hybridization assay features short analysis time, simple operation and low cost, thus has the potential to serve as the tool for PKU screening.

• Authors: Bin Chen, Xiaomian Zhou, Chunyu Li, Qiuping Wang, Dayu Liu, Bingcheng Lin
• Journal: Journal of chromatography. A (J Chromatogr A) Vol. 1218 Issue 14 Pg. 1907-12 (Apr 08 2011) ISSN: 1873-3778 [Electronic] Netherlands
• PMID: 21371716 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2011 Elsevier B.V. All rights reserved.

Chemical References

• DNA Probes
• Dimethylpolysiloxanes
• Nylons
• poly(dimethylsiloxane)-polyamide copolymer

Topics

• Compact Disks
• DNA Probes
• Dimethylpolysiloxanes
• Female
• Humans
• Microfluidic Analytical Techniques (instrumentation)
• Nucleic Acid Hybridization (methods)
• Nylons
• Phenylketonurias (blood, diagnosis)
• Pregnancy
• Prenatal Diagnosis (instrumentation, methods)
• Reproducibility of Results
• Sensitivity and Specificity
• Sequence Analysis, DNA