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INFLUENZA TYPING USING A NEW REAL-TIME DNA AMPLIFICATION ON A PORTABLE ELECTRONIC

Award Number: R44AI068537
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 01/16/2006
PERIOD OF PERFORMANCE END DATE: 02/29/2012

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INFLUENZA TYPING USING A NEW REAL-TIME DNA AMPLIFICATION ON A PORTABLE ELECTRONIC - DESCRIPTION (provided by applicant): Recent events such as the spread of swine flu and potential future detrimental mutations of influenza viruses point to an urgent need for a point-of-care influenza typing system; particularly because over 90% of hospitals and smaller clinical labs in the U.S. cannot perform DNA-based diagnostic analysis. To enable a quick response to a potential influenza outbreak, it is desirable to have a fast, accurate diagnostic method capable of simultaneously typing and subtyping influenza viruses. Innovative solutions are needed in sample preparation, DNA/RNA extraction, microfluidics, DNA amplification methods as well as instrument miniaturization to make bacterial and/or viral point-of-care molecular diagnostics a reality. This project leverages a feasibility of on-chip, real time rolling circle amplification of DNA on an electronic microarray platform demonstrated in Phase I with our efforts in miniaturization of DNA detection and development of platforms for integration with the sample preparation. The following innovative solutions form the basis of the proposed technology:(i) novel real-time fluorescence detection of (branched) rolling circle amplification of DNA on the electronic microarray platform; (ii) a disposable cartridge that incorporates a unique fluidics enabling magnetic bead DNA/RNA extraction, reverse transcription, DNA amplification and microarray detection; (iii) low cost, screen printed carbon electrodes-based microarray for electric field-assisted acceleration of DNA hybridization. The Phase II project specific aims will demonstrate the feasibility of the technology that will consist of: (1) Optimization of real-time rolling circle amplification (RCA) conditions on the carbon-based electronic microarray; (2) Development and optimization of influenza virus genotyping assay based on hemagglutinin (HA) and neuraminidase (N) typing; (3) Development of sample preparation chemistry for influenza typing assay and incorporation onto a disposable cartridge using magnetic bead based separation of RNA/DNA, reverse transcription and RCA amplification; (4) MDx system development and adaptation to influenza assay; and, (5) Validation of the influenza assay using spiked and clinical (archived) samples. PUBLIC HEALTH RELEVANCE: The recent spread of the H1N1 swine-origin influenza virus and potential detrimental mutations of influenza viruses point to an urgent need to develop point-of-care systems that will be capable of typing influenza viruses. Molecular diagnostics methods providing accurate typing of a virus will ensure a quick and appropriate response and enable improved management of patients and limit transmission of viruses. Standard techniques for viral typing are mostly based on PCR or real-time RT-PCR methods that are complex and require several hours to perform the analysis. This project brings a new real-time method for detection of DNA based on an extremely rapid rolling circle amplification of DNA/RNA targets on the electronic microarray platform. In the proposed project, the instrument and the assay will be fully developed and provide a low-cost, portable solution to diagnose influenza and/or other infectious diseases.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2011 ( Subtotal = $389,972 )
2011	2011	NEXOGEN INC	9412 MALER ROAD	SAN DIEGO	CA	92129	SAN DIEGO	USA	Allergy and Infectious Diseases Research	001	3	4/15/2011	NON-COMPETING CONTINUATION	$0
2011	2011	NEXOGEN INC	9412 MALER ROAD	SAN DIEGO	CA	92129	SAN DIEGO	USA	Allergy and Infectious Diseases Research	000	3	2/2/2011	NON-COMPETING CONTINUATION	$389,972
														Subtotal = $389,972

Issue Date FY: 2010 ( Subtotal = $398,037 )
2010	2010	NEXOGEN INC	9412 MALER ROAD	SAN DIEGO	CA	92129	SAN DIEGO	USA	Allergy and Infectious Diseases Research	000	2	3/8/2010	COMPETING CONTINUATION	$398,037
														Subtotal = $398,037

Grand Total All Awards = $788,009

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INFLUENZA TYPING USING A NEW REAL-TIME DNA AMPLIFICATION ON A PORTABLE ELECTRONIC

Award Number: R44AI068537

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 01/16/2006

PERIOD OF PERFORMANCE END DATE: 02/29/2012

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