Becton Dickinson and Company Limited

Sha-Sha Wang

Keith Edward Thornton

William A Nussbaumer

James G Nadeau

Tobin J Hellyer

Richard L Moore

James A Garrett

William E Keating

Michael Porter

Laurence Michael Vaughan

Craig C Whiteford

Feng Yang

Elaine Rose

Robert W Rosenstein

Dolores M Berger

Andrew Kuhn

David M Wolfe

Methods for predicting the development of sepsis in a subject at risk for developing sepsis are provided. In one method, features in a biomarker profile of the subject are evaluated. The subject is likely to develop sepsis if these features satisfy a particular value set. Methods for predicting the development of a stage of sepsis in a subject at risk for developing a stage of sepsis are provided. In one method, a plurality of features in a biomarker profile of the subject is evaluated. The subject is likely to have the stage of sepsis if these feature values satisfy a particular value set. Methods of diagnosing sepsis in a subject are provided. In one such method, a plurality of features in a biomarker profile of the subject is evaluated. The subject is likely to develop sepsis when the plurality of features satisfies a particular value set.

Diagnosis of sepsis

A direct chemical lysis composition includes an assay compatible buffer composition and an assay compatible surfactant. When combined with a specimen storage composition, such compositions prevent undesired modifications to nucleic acid and proteins lysed from cells in the biological sample. Assays of samples from such compositions do not require expensive and time-consuming steps such as centrifugation and prolonged high temperature processing. The direct chemical lysis composition of the present invention permits direct nucleic acid extraction from the cells in the biological sample without the need to decant off the transport media or otherwise exchange the transport media with assay compatible buffers. There is no need to combine the sample with proteinase K or another enzyme to extract nucleic acids from the cells. A method for lysing cells to obtain target nucleic acid for assay and a kit for combining the direct chemical lysis composition with a sample are also contemplated.

Methods and compositions for direct chemical lysis

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

The invention employs an unlabeled signal primer comprising a 5' adapter sequence for detection of variations in nucleic acid target sequences. The detection system further comprises a reporter probe, the 3&#x2032; end of which hybridizes to the complement of the 5&#x2032; adapter sequence of the signal primer to produce a 5&#x2032; overhang. Polymerase is used to fill in the overhang and synthesize the complement of the 5&#x2032; overhang of the reporter probe. Synthesis of the reporter probe complement is detected, either directly or indirectly, as an indication of the presence of the target.

Methods for detecting nucleic acid sequence variations

Amplification primers and methods for specific amplification and detection of a rnpB gene sequence are disclosed. The primer-target binding sequences are useful for amplification and detection of organisms of the Chlamydiaceae family in a variety of amplification and detection reactions.

Amplification and detection of organisms of the Chlamydiaceae family

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Wellesley, MA, United States of America

Sha-Sha Wang