Development of a Microfluidic Device for Detection of Pathogens in Oral Samples Using Upconverting Phosphor Technology (UPT)

Abstract:  Confirmatory detection of diseases, such as HIV and HIV‐associated pathogens in a rapid point‐of‐care (POC) diagnostic remains a goal for disease control, prevention, and therapy. If a sample could be analyzed onsite with a verified result, the individual could be counseled immediately and appropriate therapy initiated. Our group is focused on developing a microfluidic “lab‐on‐a‐chip” that will simultaneously identify antigens, antibodies, RNA, and DNA using a single oral sample. The approach has been to design individual modules for each assay that uses similar components (e.g., valves, heaters, metering chambers, mixers) installed on a polycarbonate base with a common reporter system. Assay miniaturization reduces the overall analysis time, increases accuracy by simultaneously identifying multiple targets, and enhances detector sensitivity by upconverting phosphor technology (UPT). Our microfluidic approach employs four interrelated components: (1) sample acquisition–OraSure UPlink™ collectors that pick‐up and release bacteria, soluble analytes, and viruses from an oral sample; (2) microfluidic processing–movement of microliter volumes of analyte, target analyte extraction and amplification; (3) detection of analytes using UPT particles in a lateral flow system; and (4) software for processing the results. Ultimately, the oral‐based microscale diagnostic system will detect viruses and bacteria, associated pathogen antigens and nucleic acids, and antibodies to these pathogens.

[1]  H Tanke,et al.  Use of up-converting phosphor reporters in lateral-flow assays to detect specific nucleic acid sequences: a rapid, sensitive DNA test to identify human papillomavirus type 16 infection. , 2001, Clinical chemistry.

[2]  R. Valdiserri,et al.  Twenty-five years of HIV/AIDS--United States, 1981-2006. , 2006, MMWR. Morbidity and mortality weekly report.

[3]  Amanda Paulovich,et al.  Cancer biomarkers: a systems approach , 2006, Nature Biotechnology.

[4]  T. Frieden,et al.  Applying public health principles to the HIV epidemic. , 2005, The New England journal of medicine.

[5]  Laura Kann,et al.  Youth Risk Behavior Surveillance--United States, 1993. CDC Surveillance Summaries. , 1995 .

[6]  R. Sorelle US Department of Health and Human Services gives states wider latitude in choosing services to be covered by Medicaid. , 2001, Circulation.

[7]  Shu Yang,et al.  Self-Actuated, Thermo-Responsive Hydrogel Valves for Lab on a Chip , 2005, Biomedical microdevices.

[8]  Atlanta,et al.  Revised recommendations for HIV testing of adults, adolescents, and pregnant women in health-care settings. , 2006, MMWR. Recommendations and reports : Morbidity and mortality weekly report. Recommendations and reports.

[9]  J. Eron,et al.  Acute HIV revisited: new opportunities for treatment and prevention. , 2004, The Journal of clinical investigation.

[10]  J. Gerberding,et al.  The Global HIV/AIDS pandemic, 2006. , 2006, MMWR. Morbidity and mortality weekly report.

[11]  William C Miller,et al.  Detection of acute infections during HIV testing in North Carolina. , 2005, The New England journal of medicine.

[12]  D. Malamud,et al.  Comparison of Oral Fluid Collectors for Use in a Rapid Point-of-Care Diagnostic Device , 2004, Clinical Diagnostic Laboratory Immunology.

[13]  E. Marum,et al.  A serostatus-based approach to HIV/AIDS prevention and care in Africa , 2003, The Lancet.

[14]  Mats Nilsson,et al.  Lateral-flow and up-converting phosphor reporters to detect single-stranded nucleic acids in a sandwich-hybridization assay. , 2003, Analytical biochemistry.

[15]  D. Malamud,et al.  Point Detection of Pathogens in Oral Samples , 2005, Advances in dental research.

[16]  Connie Lim,et al.  Youth risk behavior surveillance--United States, 2005. , 2006, The Journal of school health.

[17]  A. Lansky,et al.  Human immunodeficiency virus (HIV) risk, prevention, and testing behaviors--United States, National HIV Behavioral Surveillance System: men who have sex with men, November 2003-April 2005. , 2006, Morbidity and mortality weekly report. Surveillance summaries.

[18]  H. Tanke,et al.  Infrared up-converting phosphors for bioassays. , 2005, IEE proceedings. Nanobiotechnology.

[19]  J. Aberg Reconstitution of immunity against opportunistic infections in the era of potent antiretroviral therapy. , 2000, AIDS clinical review.

[20]  R. Moodie,et al.  The Global HIV/AIDS Pandemic , 2002 .

[21]  HIV prevalence among populations of men who have sex with men--Thailand, 2003 and 2005. , 2006, MMWR. Morbidity and mortality weekly report.

[22]  Shizhi Qian,et al.  Thermosiphon-based PCR reactor: experiment and modeling. , 2004, Analytical chemistry.

[23]  Zongyuan Chen,et al.  Thermally-actuated, phase change flow control for microfluidic systems. , 2005, Lab on a chip.

[24]  Michel Zuiderwijk,et al.  An amplification-free hybridization-based DNA assay to detect Streptococcus pneumoniae utilizing the up-converting phosphor technology. , 2003, Clinical biochemistry.