This project will advance the development of a point-of-care (POC) molecular diagnostic device for the rapid and inexpensive detection of infectious disease-causing pathogens in nontraditional heath care settings. An existing, fully integrated microfluidic Chemistry and Reagent Device (CARD(tm)) will be optimized for detection of pathogens in sexually transmitted infections (STIs) and urinary tract infections (UTIs). The CARD(tm) is capable of automatically isolating and lysing infectious agents, then extracting and purifying DNA and/or RNA and PCR amplifying and detecting target sequences (to date, whole blood, plasma, saliva and vaginal swabs have been successfully processed). To improve the ease-of-use of the existing CARD(tm), this project will further integrate the use of stable liposomes (at least three year real time stability has been demonstrated) to increase the signal intensity and simultaneously transduce the biological signal into an electronic one. This will be accomplished by conjugating oliogonucleotide probes that are complementary to portions of the amplified target sequences, to the exterior surface of liposomes, which, in turn, will encapsulate electroactive compounds. The resulting liposome-target complexes will be subsequently magnetically immobilized within the microfluidic channels of the CARD(tm) by use of magnetic beads conjugated to other oligonucleotide probes that are complementary to another region of the target sequences. Upon lysis of the immobilized liposomes, the released electroactive compounds will be allowed to flow onto the surface of an interdigitated ultramicroelectrode array where a continuous and reversible redox reaction will generate electric current that will be directly proportional to the concentration of the analyte. This project will therefore allow sophisticated molecular diagnostics to be performed by individuals of varying degrees of training (including unskilled), since once the sample is introduced all preparative, analytical and readout functions are automatically performed.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI082448-03
Application #
8054943
Study Section
Special Emphasis Panel (ZAI1-LR-M (J2))
Program Officer
David, Hagit S
Project Start
2009-05-15
Project End
2013-04-30
Budget Start
2011-09-15
Budget End
2012-04-30
Support Year
3
Fiscal Year
2011
Total Cost
$575,039
Indirect Cost
Name
Rheonix, Inc.
Department
Type
DUNS #
809809275
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Reinholt, Sarah J; Behrent, Arne; Greene, Cassandra et al. (2014) Isolation and amplification of mRNA within a simple microfluidic lab on a chip. Anal Chem 86:849-56
Wongkaew, Nongnoot; He, Peng; Kurth, Vanessa et al. (2013) Multi-channel PMMA microfluidic biosensor with integrated IDUAs for electrochemical detection. Anal Bioanal Chem 405:5965-74
Spizz, Gwendolyn; Young, Lincoln; Yasmin, Rubina et al. (2012) Rheonix CARD(®) Technology: An Innovative and Fully Automated Molecular Diagnostic Device. Point Care 11:42-51