Becton Dickinson and Company Limited

Millipore Corporation

Robert W Rosenstein

James G Nadeau

Song Shi

Richard Ivey

Tobin J Hellyer

Richard L Moore

Nicholas Robert Bachur, Jr

James A Garrett

Keith Edward Thornton

Dolores M Berger

Gregory Tice

Donald W Copertino

William A Nussbaumer

Thomas M Gentle, Jr

Paul Ernest Goldenbaum

Andrew Kuhn

Michael L Towns

Sha-Sha Wang

Stephen J Lovell

Gary Siuzdak

Elizabeth J Want

Zhouxin Shen

Thomas M Gentle

Timothy G Bloomster

Alan J Weiss

John K Shuler

Gene R Nathans

Leanne M Schrecengost

Sha Sha Wang

Jon C Wannlund

Abigail S Fisher

Peter Stopa

Kim P Aspden

A high-sensitivity, low-background immuno-amplification assay is provided, which offers a streamlined workflow suitable for high-throughput assays of clinically relevant samples, such as blood and other bodily fluids. The assay comprises the use of two proximity members that each comprise an analyte-specific binding component conjugated to an oligonucleotide. Binding an analyte brings the oligonucleotide moieties of the proximity members in sufficiently close contact that the oligonucleotides form an amplicon. The presence of the analyte then is detected through amplification of the amplicon and detection of the amplified nucleic acids. The sensitivity of the assay of the present invention is improved by preventing spurious or non-specific amplicon formation by proximity members that are not complexed with an analyte.

Immuno-amplification

Methods for diagnosis of HPV infection in a subject are provided. HPV infection in a subject can be determined by generating mass profile data for a biological sample from the subject and correlating the mass profile data with reference mass profiles to detect the presence or absence, and/or quantity of at least one biomarker associated with HPV infection. Methods for detecting at least one biomarker associated with HPV infection in a biological sample are also provided.

Methods for diagnosing oncogenic human papillomavirus (HPV)

The early prediction or diagnosis of sepsis advantageously allows for clinical intervention before the disease rapidly progresses beyond initial stages to the more severe stages, such as severe sepsis or septic shock, which are associated with high mortality. Early prediction or diagnosis is accomplished using a molecular diagnostics approach, involving comparing an individual's profile of biomarker expression to profiles obtained from one or more control, or reference, populations, which may include a population who develops sepsis. Recognition of features in the individual's biomarker profile that are characteristic of the onset of sepsis allows a clinician to diagnose the onset of sepsis from a bodily fluid isolated at the individual at a single point in time. The necessity of monitoring the patient over a period of time is, therefore, avoided, advantageously allowing clinical intervention before the onset of serious symptoms. Further, because the biomarker expression is assayed for its profile, identification of the particular biomarkers is unnecessary. The comparison of an individual's biomarker profile to biomarker profiles of appropriate reference populations likewise can be used to diagnose SIRS in the individual.

Diagnosis of sepsis or SIRS using biomarker profiles

Mass spectrometry techniques for determining the status of sepsis in an individual are provided. A biomarker profile resolved from a biological sample, taken from the individual, using a mass spectrometry technique is compared to a reference biomarker profile. A single such comparison classifies the individual as belonging to or not belonging to a reference population. The individual's biomarker profile and the reference biomarker profile comprise a plurality of ions each having a mass-to-charge ratio of about 100 Daltons to about 1000 Daltons. The plurality of ions can be detected by electrospray ionization mass spectrometry in positive mode. The comparison uses a decision rule, such as a classification tree, that determines the status of sepsis in the individual without requiring knowledge of the identity of the biomarkers in the biomarker profile from the individual and without requiring knowledge of the identity of the biomarkers in the reference biomarker profile.

Mass spectrometry techniques for determining the status of sepsis in an individual

Method of predicting the onset of sepsis in SIRS-positive individuals using mass spectrometry

A method for extracting ATP from a biological sample is disclosed. The method involves introducing a cationic extractant and an anionic substance and then extracting ATP. The method may be used to assay for the presence of ATP in a biological sample or to determine the amount of ATP extracted from a biological sample. The method is particularly useful in detecting contamination on surfaces and in food products. A reagent, a test device and a test kit that involve the use of the method to detect contamination are also disclosed.

ATP extraction method

The present invention relates to a method and assay device for detecting small analytes. The results of the assay can be directly read from the device, which is a lateral flow device.

Direct read lateral flow assay for small analytes

A chromatographic test strip comprising a solid support having at least a first portion and a second portion with said portions being in the same plane so as to permit capillary flow communication with each other. The sample is added to the first portion. The first portion also may comprise a tracer portion having a tracer movably supported therein. The tracer consists of a visible particulate marker. In the second portion, a binder is immobilized. The test strip is useful in a variety of immunoassays.

Solid phase chromatographic immunoassay

Test device and assay for determining analyte wherein tracer and sample may be simultaneously applied to different absorbent material portions both in capillary flow communication with an absorbent material portion having a binder supported thereon in a manner whereby sample contacts binder, prior to any substantial contact between sample and tracer or tracer and binder.

Solid phase assay employing capillary flow

I(g)G3 antibody is purified by affinity chromatography and collection of released antibody at a pH of 9.0 to 9.6. The purification is effected in a column containing both an affinity matrix and a desalting matrix, with the column being equilibrated to a pH of from 9.0 to 9.6. I(g)G3 antibody may be stored at pH 9.0 to 9.6.

Purification of antibodies

The presence of Group A Streptococcus in a biological specimen is determined from the presence of Streptococcus A antigen. A biological specimen is collected with an applicator having a plastic stick with a rayon swab. The swab is placed in an extraction reagent containing enzymes produced by the bacterium Streptomyces albus, wherein the enzymes release the antigen from the fiber. An aliquot of the extraction medium is mixed with an indicator reagent containing an antibody reactive with the antigen. The occurrence or non-occurrence of an antibody-antigen reaction is noted which indicates the presence or absence of Group A Streptococcus in the biological specimen.

Diagnostic test for Streptococcus A

A suspension of diagnostic particles comprising antibody molecules attached to a carboxylate derivatized polymer core is provided for agglutination tests. The antibody is linked to the core through an avidin-biotin bridge. Avidin is joined by an amide bond to carboxyl groups on the core, and biotin is linked by an amide bond to amino groups on the antibody molecule. The core-bound antibody is exposed to a mixture of free biotin and biotinylated antibody to attach a controlled amount of antibody that is consistent with suspension stability prior to its use in a test and rapid cross-linking of suspended particles in the presence of antigen.

Immuno-agglutination particle suspensions

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Ellicott City, MD, United States of America

Robert W Rosenstein