Eastman-kodak Company

Johnson & Johnson Clinical Diagnostics, Inc.

Ortho-clinical Diagnostics, Inc

Other

Cetus Corporation

Clinical Diagnostic Systems, Inc.

Thomas J Cummins

John Bruce Findlay

Lynn Bergmeyer

Susan Melissa Atwood

JoAnne H Kerschner

John W Sutherland

Sheryl S Sullivan

Richard C Sutton

Nancy F Green

Dennis R Zander

Ignazio S Ponticello

William Harold Donish

Tobias D Ekeze

John W H Sutherland

Fred T Oakes

Charles C Hinckley

John W Backus

Eric B Rasmussen

Ann E Casey

Paul Hong-Dze Chen

Randall D Madsen

An aqueous composition containing primers for opposing strands of two or more target nucleic acids can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of those nucleic acids. The primers for each target DNA differ in length by no more than 5 nucleotides and have a Twithin the range of from about 65 to about 74&#xb0; C., while the T's are within about 5&#xb0; C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in 'multiplexing', using multiple capture probes. All of the capture probes have T's which are greater than 50&#xb0; C. and are within 15&#xb0; C. of each other.

Diagnostic compositions, elements, methods and test kits for amplification and detection of two or more DNA's using primers having matched melting temperatures

An aqueous composition containing primers for opposing strands of human cytomegaloviral DNA and a second target DNA can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of those nucleic acids. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T within the range of from about 65 to about 74&deg; C., while the T 's are within about 5&deg; C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of hCMV DNA and other target DNA's using multiple capture probes, in &ldquo;multiplexing&rdquo;. All of the capture probes have T 's which are greater than 50&deg; C. and are within 15&deg; C. of each other.

Diagnostic compositions, elements, methods and test kits for amplification and detection of human CMV DNA using primers having matched melting temperatures

An aqueous composition containing primers for opposing strands of two or more target nucleic acids can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of those nucleic acids. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T,within the range of from about 65 to about 74&deg; C., while the T,'s are within about 5&deg; C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in &ldquo;multiplexing&rdquo;, using multiple capture probes. All of the capture probes have T,'s which are greater than 50&deg; C. and are within 15&deg; C. of each other.

Diagnostic compositions, elements, methods and test kits for amplification and detection of two or more target DNA's using primers having matched melting temperatures

The present invention relates to methods and test kits for the amplification and detection of nucleic acids from human immunodeficiency virus (HIV) type 1 and/or type 2. The methods use multiple primer sets to amplify all subtypes of HIV-1, including group M and group O isolates, and all subtypes of HIV-2.

Amplification and detection of HIV-1 and/or HIV 2

Nucleic acids can be amplified and detected using an element which has a sealable support on which is disposed a nucleic acid reagent composition. The composition is a mixture of a nucleic acid reagent composed of polymeric particles to which an oligonucleotide is covalently attached. The particles are prepared from a first polymer having a glass transition temperature of at least about 70.degree. C. and have an average diameter of from about 0.1 to about 3 micrometers. The reagent is adhered to the support using a water insoluble adhesive comprising a second polymer which has a glass transition temperature which is at least about 30.degree. C. less than the glass transition temperature of the first polymer. The adhesive is present in the composition at from about 1 to about 20 dry weight percent. The method provides high sensitivity and low background in the assay of nucleic acids, preferably using polymerase chain reaction.

Method for nucleic acid amplification and detection using adhered probes

An aqueous composition containing primers for opposing strands of two or more target nucleic acids can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of those nucleic acids. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T.sub.m within the range of from about 65.degree. to about 74.degree. C., while the T.sub.m 's are within about 5.degree. C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in 'multiplexing', using multiple capture probes. All of the capture probes have T.sub.m 's which are greater than 50.degree. C. and are within 15.degree. C. of each other.

Diagnostic compositions, elements, methods and test kits for

Diagnostic primers and probes

A target nucleic acid can be detected in a semi-quantitative fashion by passing it over detection deposits in a test element. The detection deposits include particles affixed to the test element, some of which particles have capture probe attached thereto, and other particles having no capture probe. The deposits have varying amounts of capture probe so that the signal obtained when the target nucleic acid is captured thereon can be semi-quantitatively correlated to the amount of target nucleic acid in the specimen. This method of detection can be used in nucleic acid hybridization assays or following amplification methods, including polymerase chain reaction.

Method, test element and test kit for semi-quantitative detection of

This invention provides a rapid and highly effective method for extracting nucleic acids from cells or virions without the use of proteolytic enzymes. Extraction is accomplished within a few minutes using a lysing composition comprising a buffer, a source of a DNA polymerase cofactor, a stabilizer and at least one nonionic surfactant which will release nucleic acids from cytoplasmic and nuclear membranes of cells or virions. The resulting mixture is heated to boiling for up to fifteen minutes, and the nucleic acids are recovered for amplification using polymerase chain reaction. No proteolytic enzyme is used in the extraction process.

Methods of extracting deoxyribonucleic acids without using a proteolytic

An aqueous composition containing primers for opposing strands of a target retroviral DNA (such as HIV-I DNA) can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of that target DNA and one or more additional target DNA's. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T.sub.m within the range of from about 65.degree. to about 74.degree. C., while all of the T.sub.m 's are within about 5.degree. C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in 'multiplexing', using multiple capture probes, all of which have T.sub.m 's which are greater than 50.degree. C. and within 15.degree. C. of each other.

An element has been prepared which is useful for the detection of nucleic acids in various formats. The element has a sealable support on which is disposed a nucleic acid reagent composition. The composition is a mixture of a nucleic acid reagent composed of polymeric particles to which an oligonucleotide is covalently attached. The particles are prepared from a first polymer having a glass transition temperature of at least about 70.degree. C. and have an average diameter of from about 0.1 to about 3 micrometers. The reagent is adhered to the support using a water insoluble adhesive comprising a second polymer which has a glass transition temperature which is at least about 30.degree. C. less than the glass transition temperature of the first polymer. The adhesive is present in the composition at from about 1 to about 20 weight percent. This element provides high sensitivity and low background in hybridization and other nucleic acid assays.

Element and method for nucleic acid amplification and detection using

There are described a device and method for doing confined reactions such as PCR amplification and detection, wherein solids (e.g., beads) used to obtain separation between bound and 'free' label reagents, are transferred from region to region, specifically through a wash liquid so as to wash off the 'free' label reagent from the solids. Separate chambers have dividers that are overcome by piercing or by liquification, to create passageways for the transfer of the solids. The passageways remain contained within the device.

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Rochester, NY, United States of America

Thomas J Cummins