Biotechnology Program
Center for Biotechnology and Innovation
Objective: The purpose of this initiative is the re-competition of the seven cooperative agreements (U01) awards made in September 2002 as a result of a Request for Applications (RFA) DE-02-002 (http://grants.nih.gov/grants/guide/rfa-files/RFA-DE-02-002.html) titled “Development of Technologies for Saliva/Oral Fluid Based Diagnostics”.
The current grantees possess the necessary infrastructure, organization, and leadership to finalize the integration of the individual fluidics platforms and the fabrication of portable integrated miniaturized micro-fluidic diagnostic systems able to diagnose multiple analytes of oral and systemic disease and disorders in oral fluids and address the important objectives of the validation of these diagnostic devices.
Background: To date there is a need to develop non-invasive, miniaturized, ultra-sensitive, fast detection time (~20 minutes), low cost, portable, salivary diagnostic technologies for early disease detection and to monitor individuals’ at risk of exposure to harmful biological and other agents. Traditional diagnoses have been made using blood, a relatively invasive procedure that typically requires the involvement of a health care worker. Virtually any analyte that can be detected in blood is present in human saliva in amounts that can be measured, given the appropriate instrumentation. The NIDCR, realizing the potential of saliva and oral fluids as well as the potential of new technologies and advanced instrumentation in the development of saliva and other oral fluid-based diagnostics, issued a request for applications (RFA) in January 2002 titled “Development of Technologies for Saliva/Oral Fluid Based Diagnostics”. The purpose of this RFA was to support outstanding multidisciplinary research aimed at the development of new technologies, for which proof-of-principle existed, or for the advancement of existing technologies for simultaneous multi-analyte detection in saliva and other oral fluids.
Seven awards were made in response to this RFA. They represent state-of-the-art, diverse approaches to use oral fluids for the diagnosis of oral and systemic disease. The various technologies include:
- the development and testing of micro-fluidic components such as: i) a micro-fluidic device for sample extraction; ii) saliva-based parallel diffusion immunoassay (PDIA) in a polymeric laminated disposable format; iii) inexpensive polymeric laminated devices capable of supporting surface plasmon resonance (SPRI); iv) inexpensive and simple SPRI optical system capable of imaging several surface-linked immunoassays in parallel (PSPRIIA); and test of a prototype PSPRIIA for measuring hormones in saliva
- the development of an integrated micro-fluidics platform which will contain: i) the sample separator; ii)the micro mixer needed to bring the target molecule (protein, DNA) to its probe; iii) the fluidic channel; and iv) a detection system, Light Emitting Diode (LED)
- the development of a micro-fluidic platform that will include: i) the sample collection microchip that also integrates dielectrophoresis for sample preparation and manipulation; ii) electroelution microchip iii) a sensitive microchip-based immunoassay for either cytokine profiling or oral bacteria detection ; and iv) a new epifluorescence-based miniaturized Laser-Induced Fluorescence (LIF) detector
- the development of an integrated micro-fluidic platform that can be used for the detection of a single and/or multiple pathogens. The system will contain: i) a sample collector; ii) pneumatic pump (s) for sample distribution; iii) nucleic acid extraction and a RT-PCR for amplification; iv) membranes with appropriate surface chemistries for antigen/antibody/RNA/DNA detection; and v) a detection system based on phosphors - inorganic luminescent materials, which absorb infrared energy and subsequently emit it as visible light.
- optical micro-fiber arrays for quantitative measures of a wide variety of analytes in saliva. The arrays contain different types of sensors (micro-spheres) placed into wells of an optical fiber bundle (containing up to ~25,000 wells). Detection of different analytes will be achieved by an optical imaging system
- fabrication of prototype multi-bead channel interconnected chip for the detection of cardiovascular and periodontal disease biomarkers in saliva
- a high-throughput DNA-microarray for rapid, reliable, and culture independent detection of microbial biomarkers in oral fluids as well as development of the appropriate software for interpreting microarray hybridization data.
The NIDCR realizes that each program has the ability to provide the technological advances needed for rapid, reliable, non-invasive identification of biomarker signature patterns indicative of local and systemic health status, particularly in regards to early diagnosis of diseases. The Institute also realizes the challenges these investigators are facing with regard to: i) integration of individual micro-fluidic devices into the micro-fluidic systems; ii) fabrication of micro-fluidic chips; iii) test of the automatic fluidic systems and iv) further optimization of system design
Thus, the NIDCR wishes to provide a renewal period to:
- Complete the integration of individual micro-fluidic devices
- Complete the fabrication of a portable/automated micro-fluidics-based diagnostic device
- Validate the device
- Provide feasibility analysis of large-scale (i.e., to meet estimated market demands) manufacture
- Test the device in clinical settings.
Each re-competing U01 is expected to provide a multidisciplinary structure that will coordinate activities between technology investigators, developers and clinicians in order to enable the translation of basic research to clinical application. An integral part of each re-competing application is a plan for marketing including, if necessary, plans for obtaining FDA approval.
In the first two-three years of the award each re-competing U01 will focus on finalizing and validating a portable, automated, device for simultaneous detection of analytes in health and disease (oral and/or systemic). For example, experiments should be designed to: i) integrate all the different components of the envisioned devise; ii) evaluate sensitivity, reliability and detection speed of the device; iii) verify multiple analyte detection in saliva/oral fluids; and iv) explore the potential for diagnosis of oral diseases such as oral cancer, caries, periodontal disease and systemic diseases such as cardiovascular disease, cancer, viral infections such as HIV, etc. Each U01must have two or more cross-sectional clinical studies. Manufacturing processes and clinical evaluations should be proposed in the last two years of the award period.
Current Portfolio Overview: The NIDCR extramural research portfolio, with the exception of an R21, includes only the seven U01 grants. Details of each grant are listed above.
Collaborative Activities: Because this is a re-competition of existing U01 we do not anticipate participation of other NIH Institutes.
Funding Mechanisms: The cooperative agreement (U01) mechanism will be continued in this re-competing period. Under the U01 administrative and funding mechanism the NIH assists, supports, and/or stimulates, and is substantially involved with awardees in conducting the studies by facilitating performance of the effort in a "partner" role. Details of the responsibilities, relationships, and governance of a study funded under this cooperative agreement will be the same as in the current terms and conditions of the award. Organization, as well as Terms and Condition of award remain unchanged from the original RFA (http://grants.nih.gov/grants/guide/rfa-files/RFA-DE-02-002.html). The continued use of this mechanism will allow significant programmatic input by NIDCR staff, a factor that has aided in progress during the first phase of the initiative.