SBIR-STTR Award

Real time quantitative PCR is rapidly replacing conventional or "end-point" PCR as a major diagnostic tool. Thermal Gradient Inc. (TG) has already shown that microfluidic devices based on its proprietary technology can carry out end-point PCR quickly and simply for bio-defense applications. "Quickly" is understood to mean reactions completed in minutes instead of hours, as with conventional equipment. TG would like to extend the benefits of its technology into quantitative diagnostic testing. The proposed project, Fast and Simple Real Time PCR for Diagnostic Testing, is intended to establish the feasibility of performing "real time" PCR more rapidly and conveniently than conventional means. The product envisioned here would be a compact instrument that could be used in the operating room or at the bedside to determine, among other applications, the microbial load in body fluids or the extent of tumor invasion in a biopsy, all within 30 minutes of sample collection. All of the technology required to carry this out exists today except for rapid, real-time PCR thermal cycling. In "end-point" PCR for bio-defense, the reaction is carried out to completion and then fluorescent detection is employed to identify from the amplified DNA which among many possible pathogens have been employed in an attack. In "real-time" PCR the objective is to determine from the rate of the reaction the starting concentration of usually one pathogen or a mutant sequence, although detection of multiple targets is also possible. This requires fluorescent detection of the reaction mix while it is undergoing PCR. In this Phase I application, we propose to achieve two aims: 1. Show that a "realtime" version of the TG device, one that permits observing the sample mix as it flows through the device, will still carry out an "end-point" PCR; and 2. Show that it is reasonable to expect that a well designed detection system will be able to detect the fluorescent signal that will be emitted by the "real-time" device. If these goals are met it is very likely that real time PCR can be carried out successfully in a Phase II project meant to demonstrate all the pieces of a very fast, complete, and quantitative molecular diagnostic system.

Public Health Relevance:
This Public Health Relevance is not available.

Thesaurus Terms:
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Rapid, Portable HIV Detection and Monitoring System The long term objectives of this project are: 1. Develop a prototype of a rapid portable instrument for detecting and measuring the amount of Human Immunodeficiency Viruses (HIV's) in samples of patient blood, particularly in low resource settings. 2. Carry out a pilot study to establish the effectiveness and comparative performance of the instrument against an established reference system. The aim is to provide clinicians tools to diagnose and monitor HIV that exist in resource abundant settings but are not readily available in low resource settings. In particular, the instrument system will provide clinicians with detection methods that could be helpful in identifying acute HIV-1, early diagnosis of infection in pregnant women and infants, monitoring viral load following treatment and monitoring for the emergence of antiretroviral resistance. Viral load monitoring of patients on antiretrovirals is a good example of the difference in treatment between resource abundant and low resource settings. Routinely done in the former because of the threat to patients and community should they develop a drug resistant strain of HIV, millions of recipients of antiretroviral therapy currently are not being monitored due to its cost and complexity. These aims are congruent with those parts of the mission of DAIDS "to bring an end to the HIV/AIDS epidemic by supporting the advancement of therapies for HIV infection and its complications...." This project will achieve its stated goals by carrying out a system design of a compact, fast portable system, identifying all the subsystem requirements and required technology advances and then proceeding to tackle them. The researchers at UMass Medical School will develop an effective HIV PCR assay. Thermal Gradient will carry out research on critical aspects of the key PCR amplification device and DNA sample preparation modules, and then will design and demonstrate requisite performance of the integrated test cartridge and system. Arcxis Biotechnologies is expected to contribute key DNA sample preparation technology to the integrated cartridge. Finally, the performance of the system (instrument, cartridge and assay) will be validated, first in internal tests using prepared blood samples and then a significant pilot study.

Public Health Relevance:
Rapid, Portable HIV Detection and Monitoring System A rapid HIV POC diagnostic and monitoring device would facilitate diagnosis and monitoring of antiretroviral therapy in resource-limited settings. POC tests can make HIV testing accessible in areas with limited laboratory facilities and can provide immediate test results that are needed to make clinical decisions. In particular, easy-to- use HIV tests could transform the management of pediatric HIV/AIDS in developing countries, averting millions of infant deaths, and help alleviate the threat to public health represented by the current practice of treating millions of HIV patients with antiretrovirals without monitoring their viral loads.

Public Health Relevance Statement:


Project narrative:
Rapid, Portable HIV Detection and Monitoring System A rapid HIV POC diagnostic and monitoring device would facilitate diagnosis and monitoring of antiretroviral therapy in resource-limited settings. POC tests can make HIV testing accessible in areas with limited laboratory facilities and can provide immediate test results that are needed to make clinical decisions. In particular, easy-to- use HIV tests could transform the management of pediatric HIV/AIDS in developing countries, averting millions of infant deaths, and help alleviate the threat to public health represented by the current practice of treating millions of HIV patients with antiretrovirals without monitoring their viral loads.

Project Terms:
0-11 years old; 21+ years old; AIDS Virus; AIDS test; AIDS/HIV; AIDS/HIV problem; AIDS/HIV test; ARV resistance; ARV resistant; Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; Acute; Adsorption; Adult; Antiretroviral resistance; Antiretroviral resistant; Area; Assay; Bioassay; Biocompatible Coated Materials; Biologic Assays; Biological Assay; Biotechnology; Blood; Blood Sample; Blood specimen; Child; Child Youth; Children (0-21); Clinical Evaluation; Clinical Testing; Coated Materials, Biocompatible; Communities; DNA; Deoxyribonucleic Acid; Detection; Developing Countries; Developing Nations; Devices; Diagnosis; Diagnostic; Drug resistance; Early Diagnosis; Electronics; Enzymes; Epidemic; Fluorescence; Fungi, Filamentous; Gene Products, RNA; Goals; HIV; HIV Infections; HIV test; HIV-1; HIV-1 Load; HIV-I; HIV/AIDS; HIV/AIDS problem; HIV1; HTLV-III; HTLV-III Infections; HTLV-III-LAV Infections; Human Immunodeficiency Viruses; Human T-Cell Leukemia Virus Type III; Human T-Cell Lymphotropic Virus Type III; Human T-Lymphotropic Virus Type III; Human immunodeficiency virus 1; Human immunodeficiency virus test; Human, Adult; Human, Child; Immunodeficiency Virus Type 1, Human; Infant; Infection; Injection of therapeutic agent; Injections; Investigators; LAV-HTLV-III; Laboratories; Less-Developed Countries; Less-Developed Nations; Lymphadenopathy-Associated Virus; Measures; Methods; Methods and Techniques; Methods, Other; Mission; Molds; Monitor; Operation; Operative Procedures; Operative Surgical Procedures; Optics; Output; Ownership; Patient Monitoring; Patients; Performance; Pilot Projects; Pre-Post Tests; Pregnant Women; Preparation; Public Health; Qualifying; R01 Mechanism; R01 Program; RNA; RNA, Non-Polyadenylated; RPG; RT-PCR; RTPCR; Reader; Recombinants; Reporting; Research; Research Grants; Research Personnel; Research Project Grants; Research Projects; Research Projects, R-Series; Research Resources; Research Specimen; Researchers; Resources; Reticuloendothelial System, Blood; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic Acid; SCHED; Sampling; Schedule; Specificity; Specimen; Speed; Speed (motion); Structure; Surface-Coated Materials; Surgical; Surgical Interventions; Surgical Procedure; Survey Instrument; Surveys; System; System, LOINC Axis 4; Systems Analyses; Systems Analysis; T-Lymphotropic Virus Type III Infections, Human; Target Populations; Techniques; Technology; Test Result; Testing; Thermal Conductivity; Third-World Countries; Third-World Nations; Time; Under-Developed Countries; Under-Developed Nations; Vaccines; Viral Burden; Viral Load; Viral Load result; Virus-HIV; adult human (21+); anti-retroviral resistance; anti-retroviral resistant; antiretroviral therapy; assay development; base; children; clinical decision-making; clinical test; coated materials; comparative effectiveness; cost; design; designing; drug resistant; early detection; human T cell leukemia virus III; human T lymphotropic virus III; infant death; infant measurement; infant monitoring; instrument; laboratory facility; medical schools; meetings; monitoring device; pediatric human immunodeficiency virus; pilot study; pre-clinical; preclinical; prototype; public health medicine (field); public health relevance; research clinical testing; resistance to ARV; resistance to Drug; resistance to anti-retroviral; resistance to antiretroviral; resistant strain; resistant to ARV; resistant to Drug; resistant to anti-retroviral; resistant to antiretroviral; reverse transcriptase PCR; surgery; tool; youngster