TECHNOLOGY
CASE STUDIES
Rheonix® has established worldwide corporate, governmental, and educational partnerships and has successful designed custom Rheonix Chemistry And Reagent Device (CARD®) systems for new and existing assays. Development projects include human papillomavirus (HPV) detection, warfarin dosing sensitivity and other molecular diagnostic applications. Development projects are not yet FDA cleared for human in vitro diagnostics applications.
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presented at American Association for Clinical Chemistry Annual Meeting (2010): “Fully Integrated, Automatic, and Rapid Molecular Detection and Identification of 20 Clinically Relevant HPV types using the CARD Platform”
Cervical cancer is the leading cause of cancer-related deaths among women in low-income countries and is the second leading cause of cancer-related deaths for women on a worldwide basis. Among currently FDA-approved molecular diagnostics, none are capable of distinguishing the various strains of HPV other than to classify them as high- or low-risk types.
Rheonix has developed its customized Rheonix CARD (Chemistry And Reagent Device) system capable of rapidly, easily and automatically detecting and specifically distinguishing 20 HPV types directly from clinical samples placed onto the Rheonix CARD system. A vaginal swab is collected in proprietary transport media, which allows for extended room temperature storage, if necessary. A 30 µl aliquot is applied to the Rheonix CARD sample reservoir and the run initiated. Without any further intervention by the analyst, all the following steps are automatically performed: cell lysis, nucleic acid purification, PCR amplification and multiplexed end-point detection on a low density microarray.
Rheonix conducted an analysis of 69 clinical specimens, directly comparing the diagnostic accuracy of the HPV Rheonix CARD versus a FDA-approved product. Of those specimens, 48 were found to be HPV negative by both tests, 14 were determined to be HPV positive by both tests, and 7 specimens, while negative on the comparison test, were positive on the HPV Rheonix CARD. The presence of HPV in the discordant samples was confirmed by amplicon sequencing, thus indicating the accuracy of the Rheonix CARD method. A representative microarray demonstrates the ease of detection and the presence of appropriate controls to validate proper assay performance.
This test has widespread application in both industrialized and developing nations. Rheonix has entered into a joint venture with Beijing Bohui Innovative Technology Co. to develop and manufacture a Rheonix CARD system for HPV diagnosis. Commercial launch is anticipated in 2011.
Case Study 2: Warfarin Sensitivity
Download poster presented at American Association for Clinical Chemistry Annual Meeting (2010): “CARD® Technology for Rapid and Automatic Determination of SNP Profile for Pharmacogenomics”
A significant number of life-threatening thrombolytic events are associated with the inaccurate dosing methods currently used for anti-coagulant administration. Since appropriate dosing could be faster and more safely achieved by genetic-based testing, the FDA has approved nucleic acid tests to analyze the specific single nucleotide polymorphisms (SNPs) associated with warfarin response. However, the cost and expertise required to perform these tests has been an obstacle to routine use.
Rheonix is developing a rapid, fully-integrated and automated genotyping Rheonix CARD (Chemical and Reagent Device) system for individualized dosing of Warfarin.
Use of Rheonix CARD technology significantly reduces the required skill level and the cost of equipment and materials needed to perform genotype testing. EDTA anti-coagulated whole blood is transferred to proprietary transport media that can be either stored at room temperature or immediately applied to the Rheonix CARD. Once applied to the Rheonix CARD, all required steps are automatically performed without intervention by the analyst (cell lysis, DNA purification, PCR amplification, denaturation, annealing of amplicons to filter-linked primer extension probes, primer extension in the presence of biotintylated dUTP, incubation with streptavidin-coupled HRP and color detection). Only an exact match between the immobilized primer-probe and the amplicon template will allow an extended product to be generated.
Analysis has tested and confirmed the known genotypes of commercially-available individual genomic DNAs, as well as successfully genotyping 20 individuals with previously unknown genotypes. The Rheonix CARD results were confirmed via sequencing and, in all cases, the Rheonix CARD indentified the correct genotype.
The Rheonix CARD SNP assay provides a turn-key solution to automatically perform rapid, point-of-care testing for warfarin dosing. The reduced complexity and cost of the Rheonix CARD SNP assay exemplifies how the Rheonix CARD system can be applied to an existing assay to make personalized medicine a commercially viable reality.