Eastman-kodak Company

Brent A Burdick

Susan J Danielson

Albert J Mura, Jr

Robert T Belly

Theodore W Esders

Richard C Sutton

Harold C Warren

Mark E Shafer

John Bruce Findlay

Brian A Snyder

Tai-Wing Wu

Fred T Oakes

David A Hilborn

Annie L Wu

Gregory J McClune

Robert W Zercie

Tobias D Ekeze

Ronald A Wellman

Richard W Spayd

Krishna Jayaraman

Reagents have been prepared from water-insoluble polymeric particles to which are covalently attached avidin, biotin or an avidin or biotin derivative. The polymeric particles comprise a polymer on at least the outer surface which is derived from at least one ethylenically unsaturated monomer having a reactive activated 2-substituted ethylsulfonyl, vinylsulfonyl or active halogen atom. Covalent attachment of avidin, biotin or an avidin or biotin derivative is effected either directly or indirectly through these reactive groups. The resulting reagent is useful in analytical elements and various analytical methods including agglutination and sandwich assays. The immobilized avidin, biotin or derivative can be used to complex with the corresponding biotin or avidin molecule which may be conjugated to a compound of biological interest.

Avidin-and biotin immobilized reagents and methods of use

A nucleic acid is rapidly extracted from whole blood or a peripheral blood mononuclear cell (PBMC) fraction thereof. Extraction from the PBMC fraction is accomplished by heating the fraction at or near the boiling point of water for a few minutes and recovering the extracted nucleic acid. This rapid method is particularly useful for extracting DNA for the detection of genetic diseases or infectious agents, such as HIV-I. Whole blood can likewise be heated after it is mixed with a salt solution containing a polysaccharide, such as dextran. The extracted nucleic acid is then recovered from the heated mixture. Nucleic acids extracted in this way are available for amplification using a polymerase chain reaction. Where the presence of a specific gene is to be determined for diagnostic purposes, it can be extracted as described above and subjected to suitable amplification and detection steps.

Methods of extracting, amplifying and detecting a nucleic acid from

Water soluble polymers are useful in combination with cobalt(III) redox reagents. When incorporated with the polymers, the cobalt(III) reagents exhibit improved sensitivity compared to fresh solutions of the reagents. In preferred embodiments, the cobalt(III) reagents are coated on a support using the water soluble polymer as a binder. Small areas of the coated support are then cut out and placed in a buffer solution to provide a working reagent solution. The solution is particularly useful in the testing of urine for significant bacteriuria.

Cobalt(III) reagents in combination with water soluble polymers

A method of synthesizing double-stranded DNA sequences is disclosed. The method comprises the steps of:

Method of making double-stranded DNA sequences

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

Disclosed herein is an assay useful for the quantitative determination of hemoglobin in whole blood. This assay utilizes a multizone element comprising a registration zone comprising particulate barium sulfate distributed in a 'blush' polymer, and a spreading zone having a void volume and an average pore size (e.g. greater than 5 microns) effective to accommodate whole blood. The element also contains an interactive composition capable of converting substantially all forms of hemoglobin into a detectable end product. One particularly useful interactive composition comprises a hemoglobin oxidizing agent (e.g. ferricyanide) and thiocyanate. Another useful interactive composition comprises an ionic surfactant, e.g. sodium octyl sulfate or sodium dodecyl sulfate.

Analytical element and its use in a whole blood hemoglobin assay

Reducible compounds and analytical compositions, elements and methods

Fluorescent labels comprise a polysaccharide bound to a polymeric particle which contains a fluorescent rare earth chelate. These labels can be attached to any of a variety of physiologically reactive species to provide labeled species which have improved stability in aqueous solutions. The labeled species are particularly useful in specific binding assays to determine an immunologically reactive ligand, e.g. a hapten, in either solution or dry analytical techniques.

Fluorescent labels having a polysaccharide bound to polymeric particles

A multilayer analytical element for the determination of a clinically significant enzyme analyte comprises a photosensitive compound (e.g. a photosensitive dye or dye precursor) and a filter layer containing one or more filter dyes. The filter layer is situated in the element such that incident radiation used to detect a density change resulting from interaction of the analyte and the photosensitive compound passes through the filter layer before it reaches the photosensitive compound. The use of the filter layer inhibits premature changes in the photosensitive compound caused by incident radiation. This element is particularly useful for the determination of creatine kinase or one of its isoenzymes, e.g. creatine kinase-MB.

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Rochester, NY, United States of America

Brent A Burdick