The objective of this proposal is to merge genomics, microfluidics, DNA and RNA amplification, microarrays, analytical instrumentation development and polymer microfabrication expertises to develop a revolutionary medical diagnostic device for the rapid multiparametric detection of the most relevant viruses found in respiratory tract clinical samples. Viral respiratory tract infection (VRTI) is the most common illness today (Fendrick et al., 2003), and costs over $50 billion yearly in the US alone. It is estimated that more than a dozen different viruses infect human upper airways, yet current diagnostic methods based on viral antigens often lack the required sensitivity, while culture-based methods are extremely slow. Universit? Laval with its collaborators intends to integrate their already existing diagnostic technologies into a single Microfluidic Centripetal Device (MCD), or Lab-on-a-Chip, for the detection of VRTIs (VRTI-MCD). The VRTI-MCD will automatically (i) process nasopharyngeal swab samples, (ii) extract viral RNA/DNA in a sufficiently pure form, (iii) amplify and label nucleic acids, (iv) process labelled-amplicons, (v) hybridize amplicons on low density microarrays, (vi) read and interpret the data to provide a clinically useful result within one hour, and (vii) communicate results in real-time to health information systems. The VRTI-MCD will be a disposable polymer-based device that will encase all necessary reagents. The disposable will be processed by a portable and easy-to-use opto-mechanical instrument enabling automated VRTI diagnostics. Seventeen viruses, including influenza A (such as avian flu and swine flu), influenza B, SARS coronavirus, parainfluenza, and adenovirus responsible for the majority of VRTI along with key antiviral resistance markers will be detectable using a single VRTI-MCD. This novel diagnostic platform will be developed and optimized using nasopharyngeal swab specimens available in the extensive clinical samples collection at the Centre de Recherch? en Infectiologie (CRI). By the end of year-five support by NIH, we will have validated the integrated VRTI-MCD using human clinical samples and the portable instrument will be ready for worldwide clinical testing for diagnostics at the point-of-care. Thereafter, within two years, product development and approval by health authorities (FDA, Health Canada, etc.) will be undertaken by a commercial partner. This new technological platform will result in an unprecedented expansion of point-of-care diagnostic tests (POCTs) for viral respiratory tract infections, while providing rapid yet economical molecular diagnostics results in less than one hour instead of the current 2-3 days. This will improve medical response, allowing for the timely use of proper antiviral drugs and to ensure that rapid quarantine measures only be used when appropriate. These POCTs will help public health authorities to define new operational strategies for both lab-based and field-based applications.
Our transdisciplinary collaborative team will develop an adaptable, rapid, sensitive, specific, and fully automated multiparametric biodefense diagnostic device for use at the point-of-care. The device will target 17 viruses responsible for respiratory tract infections, from which 5 are included in NIAID Category C viral pathogens, namely influenza A, influenza B, avian flu (H5N1), swine flu (H1N1), viruses and the SARS coronavirus. The proposed platform will allow quicker patient management and optimal treatment when necessary, thus reducing the contagion rate, a very significant advantage especially in the case of a pandemic. It will also prevent the inefficient and over prescription of anti-bacterial agents (it is estimated that close to 90% of patients with a diagnosis of influenza receive antibiotics), thereby reducing antibiotic resistance, a major burden in healthcare systems worldwide. Moreover, the availability of virus-specific diagnostic tools will favour the development of specific antiviral drugs by pharmaceutical companies.
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