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A Rapid, Self-confirming Assay for HIV: Simultaneous Detection of Anti-HIV Antibodies and Viral RNA
- Source :
- Journal of AIDS & clinical research
- Publication Year :
- 2016
-
Abstract
- Objective: We developed a microfluidic system to simultaneously detect host anti-HIV antibodies and viral RNA in the same specimen in order to satisfy two important diagnostic criteria, especially within resource-limited settings. First, the system can detect acute HIV infection and allow immediate confirmation of a seropositive screening result by detection of HIV RNA. It also addresses the well-known "seroconversion window" during early HIV infection when antibodies are not yet detectable and viral loads are at their highest. Methods: We first developed and optimized two separate manual assays for the detection of host anti-HIV antibodies and viral RNA and then converted them to the microfluidic system. We optimized a commercially available serologic assay to run within the microfluidic device while we incorporated the isothermal LAMP assay to detect the presence of viral RNA. The microfluidic device and instrumentation were developed to simultaneously perform both assays without any user intervention. Results: The finalized system consists of a disposable injection molded and film-laminated microfluidic CARD disposable device and a portable, software controlled instrument, which together can automatically perform all steps of both assays without any user intervention after the initial loading of samples and reagents. The microfluidic CARD cartridge has multiple microchannels, valves, pumps and reservoirs, which perform the immunoassay, isolates viral RNA for detection by magnetic bead based purification, and Reverse Transcriptase loop-mediated isothermal amplification (RT-LAMP). The microfluidic system was able to detect host anti-HIV antibodies and viral RNA in either a blood or saliva sample. Conclusion: The ability to detect antibodies and simultaneously confirm a seropositive HIV-RNA result provides healthcare workers with a complete and accurate appraisal of a patient's infection status in the earliest stages of the disease and represents an important tool for the "Test and Treat" and "Treatment as Prevention" approaches for controlling the HIV epidemic.
- Subjects :
- 0301 basic medicine
Immunology
Loop-mediated isothermal amplification
Dermatology
Article
Serology
03 medical and health sciences
Virology
medicine
Molecular diagnostics
Seroconversion
biology
medicine.diagnostic_test
business.industry
HIV diagnostics
Rapid diagnostics
Reverse transcriptase
3. Good health
AIDS
030104 developmental biology
Infectious Diseases
Immunoassay
biology.protein
Antibody
business
Viral load
Loop Mediated Isothermal Amplification (LAMP)
Subjects
Details
- Language :
- English
- ISSN :
- 21556113
- Volume :
- 7
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of AIDS & clinical research
- Accession number :
- edsair.doi.dedup.....356c2ff09be132d261a80da57839f17e