Microchip-based system for hiv diagnostics
OwnerBruce Walker, William Rodriguez +1inventors
InventorsBruce Walker, William Rodriguez, John McDevitt
Life of this patent
Estimating the term…
Abstract
The invention relates to microchip-based assays to measure HIV-associated analytes of interest (e.g., CD4 lymphocytes, HIV RNA and liver enzymes) in a sample from a subject infected with the HIV virus. Methods of the present invention are optimal for use in monitoring HIV disease in resource-poor settings.
Claims30
A method for detecting an HIV-associated analyte in a sample of blood comprising:
- passing an HIV-associated analyte coupled to a fluorescence-emitting conjugate through a filter, wherein the analyte is immobilized on the filter thereby separating the analyte from the blood;
- delivering light to the analyte at a wavelength suitable for excitation of the conjugate;
- digitally imaging a conjugate emitted fluorescent signal using a detector.
(canceled)
7-11. (canceled)
14-15. (canceled)
A method for determining the ratio of CD4+ T cells to CD8+ T cells in a sample of blood comprising:
- passing CD4+ and CD8+ T lymphocytes through a filter, wherein the CD4+ and a CD8+ T lymphocytes are captured on the filter;
- coupling the CD4+ and the CD8+lymphocytes to different fluorescence emitting conjugates;
- delivering light to the lymphocytes at a wavelength suitable for excitation of each conjugate;
- digitally imaging each conjugate emitted fluorescent signal using a detector;
- and
- determining the ratio of CD4+ cells to CD8+ cells.
A method for detecting HIV-RNA in a sample of blood comprising:
- passing a blood sample comprising HIV-RNA through a flow cell comprising one or more cavities, wherein one or more of the cavities contains a filter comprising a complementary nucleotide sequence coupled to a fluorescence-emitting compound;
- forming a binding complex between the HIV-RNA and the complementary nucleotide sequence on the filter, whereby the signal emitted from the fluorescence-emitting compound is enhanced by formation of the binding complex;
- delivering light to the complex at a wavelength suitable for excitation of the fluorescence-emitting compound;
- and
- digitally imaging a fluorescent signal emitted by the complex using a detector.
A method for detecting an HIV-associated analyte in a sample of blood comprising:
- passing an HIV-associated analyte coupled to a fluorescence-emitting conjugate through a flow cell comprising one or more cavities, wherein one or more of the cavities contains an agarose bead comprising a suitable biological agent having binding affinity for the analyte, such that the analyte is immobilized on the bead, thereby separating the analyte from the blood;
- delivering light to the analyte at a wavelength suitable for excitation of the conjugate;
- digitally imaging a conjugate emitted fluorescent signal using a detector.
(canceled)
25-26. (canceled)
28-29. (canceled)
32-33. (canceled)
The method of claim 33, wherein the fluorescence-emitting conjugate comprises an anti-CD3 antibody, and wherein the fluorescence-emitting conjugate is an Alexa647 conjugated anti-CD3 antibody.
A method for determining the ratio of CD4+ T cells to CD8+ T cells in a sample of blood comprising:
- passing CD4+ and CD8+ T lymphocytes, each coupled to a different fluorescence-emitting conjugate, through a flow cell comprising one or more cavities, wherein one or more of the cavities contains an agarose bead comprising a biological agent having binding affinity for either CD4+ or CD8+ T lymphocytes, such that the lymphocytes are immobilized on the bead, thereby separating the analyte from the blood;
- delivering light to the lymphocytes at a wavelength suitable for excitation of each conjugate;
- digitally imaging each conjugate emitted fluorescent signal using a detector;
- and
- determining the ratio of CD4+ cells to CD8+ cells.
A method for detecting HIV-RNA in a sample of blood comprising:
- passing a blood sample comprising HIV-RNA through a flow cell comprising one or more cavities, wherein one or more of the cavities contains an agarose bead comprising a complementary nucleotide sequence coupled to a fluorescence-emitting compound;
- forming a binding complex between the HIV-RNA and the complementary nucleotide sequence on the bead, whereby the signal emitted from the fluorescence-emitting compound is enhanced by binding of the HIV-RNA to the complementary nucleotide sequence;
- delivering light to the complex at a wavelength suitable for excitation of the fluorescence-emitting compound;
- and
- digitally imaging a fluorescent signal emitted by the complex using a detector.
Dependencies are inferred from the claim text.