Leland Stanford Junior University

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Ronald L Rabin

Leonore A Herzenberg

Leonard A Herzenberg

Mario Roederer

William E Benitz

Viraj Pramod Mane

Erica E Weirich

Yvonne Maldonado

The invention provides highly sensitive, specific and efficient quantitative real-time PCR compositions, methods and assay kits to detect at least one IFN subtype and/or IFN subtype allotypic variants. Primer/probe sets complementary to the coding sequence of an IFN subtype of interest avoid spurious detection of degraded mRNA and enhances the correlation between the IFN subtype that is measured by the assays of the invention and the protein that is actually expressed. The invention also provides methods for designing primers and methods of using the compositions and assay kits. The compositions, kits, and methods of the invention may be used, for example, to monitor vaccine efficacy, autoimmune disease, chronic infections, or tumor therapy.

Compositions for detecting human interferon-alpha subtypes and methods of use

A rapid assay for infection in immunodeficient patients such as neonates or immunocompromised patients (e.g. HIV or transplant patients) allows diagnosis at initial evaluation, such that antibiotic treatment and confinement to an intensive care unit can be avoided for uninfected patients. The assay can be used for detecting bacterial, viral, or fungal colonization of the blood stream, cerebrospinal fluid (CSF), or urinary tract. The method is particularly useful for sepsis diagnosis. Polymorphonuclear leukocyte (PMN, neutrophil) CD11b (Mac-1, CR3) levels are measured by flow cytometry or laser scanning microscopy in low volume (0.1 ml) whole blood samples. A dual-laser FACS identifies neutrophils by FITC-conjugated anti-CD15 fluorescent antibodies, and identifies surface neutrophil CD11b marked with PE-conjugated anti-CD11b antibodies. Spontaneous upregulation of CD11b is prevented by handling samples at 4.degree. C. or adding a stabilizing compound such as anti-CD14 antibody or adenosine to the samples. A kit comprises calibration markers such as CD11b-coated microspheres for calibrating the device used for analysis according to diagnosis threshold(s).

Rapid assay for infection in neonates

A method of identifying an abnormal T-cell profile of an immunocompromised subject is disclosed. The method involves detecting the immunoreactivity of T-cells with at least two antibodies selectively reactive with naive T-cell surface proteins and comparing data from cells isolated from an immunocompromised subject with data from cells isolated form a non-immunocompromised subject. Also disclosed are methods of screening drugs or compounds for efficacy to stimulate the production of naive T cells in a subject.

Methods for determining T-cell profiles of immunocompromised subjects

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Rockville, MD, United States of America

Ronald L Rabin