title: Integration of clinical point-of-care requirements in a DNA microarray genotyping test.
abstract: Various proof-of-concept studies have shown the potential of biosensors with a high multiplex detection capability for the readout of DNA microarrays in a lab-on-a-chip. This is particularly interesting for the development of point-of-care genotyping tests, to screen for multiple pathogens and/or antibiotic resistance patterns. In this paper, an assay workflow is presented, suited for the development of novel lab-on-a-chips with an integrated DNA microarray. Besides the description of the different assay steps (DNA purification, amplification and detection), a control strategy is presented according to recommendations of the US Food and Drug Administration (FDA). To use a lab-on-a-chip for diagnostic applications, the optimization and evaluation of the assay performance with clinical samples is very important. Therefore, appropriate quantification methods are described, which allow optimization and evaluation of the separate assay steps, as well as total assay performance. In order to demonstrate and evaluate the total workflow, blood samples spiked with Streptococcus pneumoniae were tested. All blood samples with ≥ 10(3)CFU S. pneumoniae per ml of human blood were successfully detected by this genotyping assay.
authors: Janssens Diagnostics, Turnhoutseweg 30, 2340 Beerse, Belgium. Electronic address: bvandors@its.jnj.com.
authors: Bieke
authors: B
authors: Van Dorst
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Centre, Nijmegen, The Netherlands; Laboratory of Medical Immunology, Department of Laboratory Medicine, Radboud University Medical Centre, Nijmegen, The Netherlands.
authors: Amelieke
authors: A
authors: Cremers
authors: IMEC, Leuven, Belgium.
authors: Karolien
authors: K
authors: Jans
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Centre, Nijmegen, The Netherlands; Laboratory of Medical Immunology, Department of Laboratory Medicine, Radboud University Medical Centre, Nijmegen, The Netherlands.
authors: Trees
authors: T
authors: Van Domburg
authors: Janssens Diagnostics, Turnhoutseweg 30, 2340 Beerse, Belgium.
authors: Kim
authors: K
authors: Steegen
authors: IMEC, Leuven, Belgium.
authors: Chengjun
authors: C
authors: Huang
authors: Robert Bosch GmbH, Gerlingen, Germany.
authors: Christian
authors: C
authors: Dorrer
authors: IMEC, Leuven, Belgium; Department of Physics and Astronomy, Catholic University of Leuven, Celestrijnenlaan, 200d, 3001 Herverlee, Belgium.
authors: Liesbet
authors: L
authors: Lagae
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Centre, Nijmegen, The Netherlands; Laboratory of Medical Immunology, Department of Laboratory Medicine, Radboud University Medical Centre, Nijmegen, The Netherlands.
authors: Gerben
authors: G
authors: Ferwerda
authors: Janssens Diagnostics, Turnhoutseweg 30, 2340 Beerse, Belgium.
authors: Lieven J
authors: LJ
authors: Stuyver
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation; D004269:DNA, Bacterial / Q000235:genetics; D004867:Equipment Design; D060005:Genotyping Techniques / Q000295:instrumentation*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D011008:Pneumococcal Infections / Q000097:blood / Q000175:diagnosis; D019095:Point-of-Care Systems; D012680:Sensitivity and Specificity; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial
keywords: DNA hybridization; Flow-through; Genotyping assay; Gold; Lab-on-a-chip; Spiked blood samples