Eastman-kodak Company

Ortho-clinical Diagnostics, Inc

Johnson & Johnson Clinical Diagnostics, Inc.

Other

Veridex, Lcc

Clinical Diagnostic Systems, Inc.

Robert T Belly

Albert J Mura, Jr

Vanessa R Lum

Patricia M Scensny

Peggy W Cichanowicz

Theodore W Esders

Bruce E Babb

Brent A Burdick

David S Frank

Jianbo Sun

Michele McWilliams Steinmann

Robert F Guardino

Eric R Schmittou

Paul B Gilman, Jr

Drake M Michno

Brian A Snyder

John B Chemelli

Annie L Wu

Lee J Fleckenstein

Maurice A Kildal

Sheryl S Sullivan

William N Washburn

Thaddeus D Koszelak

Chung-yuan Chen

Martha Miller Kopcienski

Caroline Erdrich

Seymour Zigman

Jacqueline Toner

Gary J Chilson

John Backus

John Robert Chemelli

Laurie J Clements

Nucleic acids can be made available for amplification or other treatment after admixture of a sample with specific weakly basic polymers to form a precipitate with the nucleic acids at acidic pH. After removing non-precipitated materials, the pH is then made basic, thereby releasing the nucleic acids from the polymer. This method for preparing specimen samples is simple and quite rapid, and the released nucleic acids can be further treated in hybridization assays or amplification procedures. No surfactant or other cell lysing reagents are employed. The weakly basic polymers are water-soluble and cationic at acidic pH, but neutral in charge at basic pH.

Rapid and efficient capture of DNA from sample without using cell lysing reagent

A method of extracting RNA from biological systems (cells, cell fragments, organelles, tissues, organs, or organisms) is presented in which a solution containing RNA is contacted with a substrate to which RNA can bind. RNA is withdrawn from the substrate by applying negative pressure. No centrifugation step exceeds thirty seconds. Preferably, the RNA is diluted prior to filtration and one or more wash steps can be used to remove interferents. In one such method, except for DNA shearing and drying steps, centrifugation is not undertaken during the extraction step.

Rapid extraction of RNA from cells and tissues

The present invention provides rapid and highly effective methods for extracting nucleic acids suitable for PCR amplification from tissue samples and paraffin-embedded tissue samples. Extraction is accomplished within a few minutes using a composition comprising a buffer, at least one nonionic surfactant, and a protease enzyme. The sample is then heated at alkaline pH and after a centrifugation step, the DNA is the supernatant can be used directly in known amplification methods, such as PCR.

Methods for extracting nucleic acids from tissue samples and paraffin-embedded tissues

The present invention provides a novel dry analytical element for determination of an analyte in a fluid sample in thin film immunoassay, particularly chemiluminescence, fluorescence and colorimetry immunoassay. There is provided an analytical element comprising a support member, a reagent matrix comprised of at least one layer of a porous material carried on the support member, the matrix including a read area, an absorbent material in contact with the matrix, and light absorbing or reflecting means integral to the absorbent material and/or positioned between the absorbant material and the read area for absorbing or blocking background signal generated by washed, removed label and other interferents from outside the read area prior to it moving back into the read area. The analytical element can comprises a cover depending on the type of immunoassay to be conducted.

Format for immunoassay in thin film

A flexible container containing pre-deposited reagents, comprising a plurality of temporarily sealed, breakable compartments dispersed in the container, each compartment containing a reagent useful for immuno-assaying, and each compartment comprising opposed confining walls at least one of which is sufficiently flexible as to allow the compartment to be compressed in the presence of an adequate external force, external outlets in the container for the contents of said compartments, passageways extending from each of the compartments to the outlets, each of the outlets terminating in a platform that supports a drop of at least 10 .mu.L as a pendant drop, the platforms of at least two of the outlets being adjacent so as to form between them an exterior angle with respect to each other sufficient to cause drops pendant therefrom to substantially uniformly intermix.

Container for staining of cells and tissues in combination with a roller

A slide for reacting a biological sample with reagents, and a method for using the same. The slide comprises a liquid-impervious top surface comprising glass, first portions of the surface having thereon a hydrophobic mask that repels an aqueous liquid, and second portions of the surface being completely free of any coverage by the mask so as to be wettable with an aqueous liquid, at least one of the other wettable second portions being totally isolated from the other wettable second portions by some of the mask, so that aqueous liquid will not flow from the one wettable portion to the another wettable portion until an overflow amount of aqueous liquid is added to the one wettable portion in an amount sufficient to bridge across the mask, and an absorbent material mounted at one side of the surface in position to receive liquid from the slide. The method of use features confining aqueous reagents to the area restricted by the hydrophobic mask, except for when additional amounts are added to force at least some of the liquid to move towards a side edge of the slide, where most of the liquid is absorbed.

Apparatus for staining of cells and tissues

Certain water-compatible reducible compounds are useful in analytical compositions and elements for assay of various analytes, e.g. microorganisms. These compounds comprise a moiety which provides a detectable species (e.g. a dye) when released from the compound at physiological pH. Further, these compounds are aromatic derivatives or quinones having water-compatibilizing substituents which allow them to be used in compositions without the use of surfactants.

Method for providing chemically or biologically useful moiety from

Compounds and reagents containing the compounds are disclosed for detecting oxidase positive organisms. The compounds have the structure

Reagents for detecting oxidase positive microorganisms

Certain reducible compounds are useful in analytical compositions, elements and methods, e.g. for assays of bacterial cells. These compounds comprise a moiety which provides a detectable species (e.g. a dye) when released from the compound in an environment of pH 9 or less (i.e. physiological pH). Structurally, the reducible compounds are aromatic derivatives or quinones having suitable substituents which promote varying amounts of moiety release at physiological pH. When reduced at about pH 7, the preferred compounds release at least 50% of the available detectable species within 30 minutes.

Analytical methods utilizing reducible components

Certain water-compatible reducible compounds are useful in analytical compositions and elements for assay of various analytes, e.g. microorganisms. These compounds comprise a moiety which provides a detectable species (e.g. a dye) when released from the compound at physiological pH. Further, these compounds are aromatic derivatives or quinones having water-compatibilizing substituent which allow them to be used in compositions without the use of surfactants.

Water-compatible reducible compounds and their use in analytical

Hydrolyzable substrates comprise blocked moieties which, when cleaved from the substrate during hydrolysis, provide electron transfer agents. The released electron transfer agents can be recycled between a reductant and a reducible compound that upon reduction provides a detectable species. Alternatively, they can be recycled between an oxidant and an oxidizable compound that upon oxidation provides a detectable species. These substrates are useful in analytical compositions, elements and methods for the determination of hydrolytic analytes, such as hydrolytic enzymes or biological cells containing such enzymes.

Hydrolyzable compounds which release electron transfer agents and

A method for the determination of cells in a sample is disclosed. The sample is typically urine and the method is useful in removing interfering reductants from the sample. The method comprises the steps of:

Method for minimizing interference by reductants when detecting cells in

Rapid differentiation between viable gram-positive and gram-negative bacteria is accomplished with certain polypeptide antibiotics which are used in combination with a compound which is normally reducible by the bacteria. The antibiotics selectively inhibit the reduction of the reducible compound by gram-positive bacteria but do not substantially affect the reducing capacity of the gram-negative bacteria. The particular antibiotics useful are cyclic polypeptides which affect the function of the cytoplasmic membrane of bacteria. A particular polypeptide antibiotic, polymixin B, will distinguish Proteus bacteria from other gram-negative genera.

Rapid differentiation of bacteria using cyclic polypeptide antibiotics

Rapid differentiation between viable fungi (for example, yeast) and viable bacteria is accomplished with certain polyene antibiotics which are used in combination with a compound which is normally reducible by both the fungi and the bacteria. The antibiotics selectively and substantially inhibit the reduction of the reducible compound by the fungi but do not affect the reducing capacity of the bacteria. The particular antibiotics useful are polyenes which selectively affect the function of the cytoplasmic membrane of fungi.

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Webster, NY, United States of America

Robert T Belly