Integration of clinical point-of-care requirements in a DNA microarray genotyping test

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 ≥103CFU S. pneumoniae per ml of human blood were successfully detected by this genotyping assay. •An assay workflow is presented that can be used for the development of a lab-on-a-chip test with an integrated DNA microarray.•A control strategy is described according to the requirements of the FDA to allow the use of this assay for diagnostic application.•Quantification methods are described to validate assay’s performance with clinical samples.•The assay workflow and quantification methods were demonstrated with spiked blood samples.