Becton Dickinson and Company Limited

Arbor Vita Corporation

Adrien P Malick

John D Mantlo

Hans H Feindt

Susan M Kircher

William B Brasso

Liping Feng

Vanda White

Julie L Rosales

Song Shi

Stephen C Wardlaw

Robert A Levine

Rodolfo R Rodriguez

Ben Turng

Gerald D Hahn

Alvydas J Ozinskas

Jon E Salomon

Xiao Mo

Dyan Luper

Tuan-Linh Ngoc Nguyen

Leon W Terstappen

Patrick Shawn Beaty

Stephen J Lovell

Edward C McFarland

Virginia M Crews

Kristen L Manion

Curtis M Gosnell

James Y Zhou

Donald Ray Callihan

Subhash Dhanesar

Jeffery H Bruton

Carrie S Ferguson

Jeff H Bruton

Robert J Beadenkopf

John P Douglass

Gretta Campbell

Jefferey H Bruton

Donald R Callihan

Shawn Beaty

Various embodiments disclosed herein provide for reagents and methods for rapidly isolating viable microbial cells, including, from positive blood culture samples. The resulting microbial pellet can be used for both identification and growth-based methods such as antimicrobial susceptibility testing. The buffers described herein may contain a base solution, non-ionic detergents, thiols, and optionally, ammonium chloride. The disclosed methods provide a process for rapidly isolating and concentrating viable microorganism(s) from PBC samples using only one sample preparation tube and centrifugation while removing cellular debris from the mammalian blood cells that may interfere with identification methods.

Formulations and process for isolating viable microorganisms from positive blood cultures

Rapid methods that identify sepsis-causing bacteria or yeast aid the physician in critical therapeutic decision-making, thus decreasing patient mortality rates. The methods described herein employ plating microorganisms directly on to a MALDI-MS plate, adding concentrated formic acid, and identifying the microorganism by mass spectrometry. Optionally, an organic solvent may be combined with the formic acid, or added to the sample before or after the concentrated formic acid is added thereto. The methods enable direct extraction of proteins from microorganisms without the need for liquid protein extraction methods and yields positive identification results for gram-positive bacteria, gram-negative bacteria and yeast in minutes.

Identification of microorganisms using MALDI-TOF-MS on-plate extraction

Various embodiments disclosed herein provide for reagents and methods for rapidly isolating viable microbial cells, including, from positive blood culture samples. The resulting microbial pellet can be used for both identification and growth-based methods such as antimicrobial susceptibility testing. The buffers described herein may contain a base solution, non-ionic detergents, thiols, and optionally, ammonium chloride. The disclosed methods provide a process for rapidly isolating and concentrating viable microorganism (s) from PBC samples using only one sample preparation tube and centrifugation while removing cellular debris from the mammalian blood cells that may interfere with identification methods.

Methods for producing a protein extract from cells, such as cells or cellular samples containing viral proteins, are provided. In general terms, the methods may involve: increasing the pH of the cells to a pH of at least about pH 10.0 to produce an intermediate composition, and then, in the presence of a non-ionic detergent such as a polyoxyethylene alkyl ether, neutralizing the pH of the intermediate composition to produce the protein extract. Such methods can be used in conjunction with methods for detecting one or more target proteins in a sample, such as viral proteins. Systems, kits and compositions for practicing the subject methods are also provided.

Method of efficient extraction of protein from cells

Methods of the invention include the isolation of intact, viable microorganism(s) from positive blood culture ('PBC') samples for use in downstream analyses such as identification and antimicrobial susceptibility testing ('AST'). The methods involve collecting a portion of the PBC sample, adding a choline-containing solution, lysing the blood cells, isolating the viable microorganism, and performing downstream analysis of the isolated, viable microorganism. The methods can be applied to a variety of gram-positive bacteria, gram-negative bacteria, and/or yeast, and particularly to strains of

Method for direct and rapid identification of microorganisms and antimicrobial susceptibility testing from positive blood cultures

The present invention is directed to non-instrumented assays giving quantitative and/or qualitative results. In one embodiment, the present invention is directed to a process and kit determining analyte in a sample wherein the solid support has a contact zone and binder distributed and immobilized throughout the solid support. A sample and a tracer are added to the contact zone and a visible zone is obtained. In another embodiment, the present invention is directed to a process and test wherein the solid support has two contact zones and the binder is not immobilized on the solid support. In this embodiment, the binder is mobile and is added to the second contact zone. At the same time, sample and a tracer are added to the first contact zone. A visible zone is obtained.

Non-instrumented assay with quantitative and qualitative results

The invention is directed to a non-instrumented assay giving quantitative and/or qualitative results. Specifically, the invention is directed to an assaying process wherein a first portion of binder supported on substantially the entire length of a solid is contacted with a sample suspected of containing analyte. The binder is a binder for at least the analyte. The first portion is also contacted with a tracer. The process includes contacting a second binder portion also supported on the solid with tracer. The second portion is juxtaposed to the first portion and has a known amount of analyte thereon. The amount of analyte in the sample is determined by comparing the visibility of tracer in the first and second portions. In a preferred embodiment, a third binder portion supported on the solid juxtaposed to the second portion is contacted with tracer. The third portion has a known amount of analyte thereon. The amount of analyte in the third portion is different from the amount on the second portion. According to another aspect, the invention is also directed to a kit for performing the above-described process.

A patient's health may be diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more bodies or groups of bodies such as floats, inserts, liposomes, or plastic beads of different densities. Each density-defined body carries analyte-capture binding materials such as antigens or antibodies, which are specific to an epitope, or other specific high affinity binding site on a target analyte which target analyte may be in the blood or other sample being tested; and the level of which analyte is indicative of the patient's health. At least one labeled binding material which is also specific to an epitope, or other specific high affinity binding site on the target analyte is added to the sample so as to form labeled binding material/analyte/body complexes in the sample. Upon centrifugation, the complexes will settle out in different areas in the tube according to the respective density of the body or bodies; and the degree of label emission of the complex layers can enable qualitative and/or quantitative analyses of the sample to be made. Unbound labeled binding materials will be separated from the complexed layers by the washing action of ascending or descending components of the sample during the centrifugation step. Unbound labeled binding material will thus not interfere with the analysis.

Assay of blood or other biologic samples for target analytes

Stabilized microspherical particles having hydrophobic liquid cores prepared as oil-in-water microemulsions. The particles are stabilized by a surface layer comprising an amphiphilic compound and may be functionalized to allow covalent coupling of a ligand to the surface of the particle. When used as tracers in assays, a water insoluble dye may be incorporated in the core liquid of the microparticles.

Stabilized microspheres and methods of preparation

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Granite, MD, United States of America

Adrien P Malick