Eastman-kodak Company

Johnson & Johnson Clinical Diagnostics, Inc.

Other

Ortho-clinical Diagnostics, Inc

Clinical Diagnostic Systems, Inc.

John Bruce Findlay

Thomas J Cummins

Lynn Bergmeyer

Susan Melissa Atwood

JoAnne H Kerschner

John W Sutherland

Ignazio S Ponticello

Richard C Sutton

William Harold Donish

Harold C Warren

Fred T Oakes

Marlene M King

Susan J Danielson

Annie L Wu

Marsha D Oenick

Charles C Hinckley

John W Backus

Jeffrey A Wellman

Paul Nicholas Schnipelsky

Leonard J Seaberg

John W H Sutherland

Dennis R Zander

Mark E Shafer

Theodore W Esders

Corey H Levenson

Chu-an Chang

Brent A Burdick

Gary E Norton

Tobias E Ekeze

Shirley Y Lynn

Richard M Schubert

Jerome C Swartz

Ronald A Wellman

Paul Hong-Dze Chen

Paul H-d Chen

Janice M Mayer

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

A cuvette and a method of use prevent nucleic acid amplified by PCR technology from being released to the atmosphere, while still proceeding to a detection step to determine whether or not the nucleic acid is present. Detection reagents are either pre-incorporated into compartments in the cuvette or added after amplification. In the latter case, a check valve prevents amplified nucleic acid from being released. Transfer of liquids between compartments is achieved via the use of flexible compartment walls and an external pressure source, or via pistons that are part of the cuvette and operate on the compartments as a piston within a piston chamber.

Containment cuvette for PCR and method of use

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

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

Nucleic acids can be amplified and detected using a very rapid polymerase chain reaction procedure in which two different nucleic acid sequences are present. This method allows one to preferentially modulate (for example, suppress) the degree of amplification of one or more nucleic acid sequences relative to other nucleic acid sequences. This modulation is achieved by exploiting differences in the relative primer melt temperatures, or by using certain ratios of primers. Each PCR cycle is very fast, that is less than about 90 seconds. This method is particularly useful for amplification and detection of DNA associated with infectious agents that may be present in a specimen in very small quantities compared to other nontargeted nucleic acids.

Rapid method for preferential coamplification of two different nucleic

Diagnostic primers and probes

Nucleic acids can be made available for amplification or other treatment after lysis by contacting the lysate 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. The weakly basic polymers are water-soluble and cationic at acidic pH, but neutral in charge at basic pH.

Methods for capture and selective release of nucleic acids using weakly

A nucleic acid test article can be used to detect a targeted nucleic acid found in a specimen. The test article includes a substrate having two opposing surfaces and a water-insoluble nucleic acid probe attached in a distinct zone of one of the surfaces. The probe is prepared from a water-insoluble particle to which is covalently attached an oligonucleotide which is complementary to the targeted nucleic acid. Substantially none of the probe is embedded within the surface of the substrate. Particularly useful test articles have a multiplicity of water-insoluble probes located in distinct zones on one of the substrate surfaces. These probes are useful for the detection of a multiplicity of targeted nucleic acids, particularly after amplification by polymerase chain reaction.

Nucleic acid test article and its use to detect a predetermined nucleic

A device and method are disclosed for amplifying and detecting nucleic acid material. The device and method use a label and signalling material responsive to the label to produce a detectable signal. A surprising result of the method and device is that at least one of the wash steps heretofore required has been eliminated without substantially adversely affecting the results.

Nucleic acid material amplification and detection without washing

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 oligonucleotide is linked to a particle through a protein or carbohydrate to form a water-insoluble nucleic acid probe. The protein or carbohydrate has a pI of about 6 or less, and has been chemically modified with an acylating, alkylating or sulfonylating agent. The particle surface is substantially free of other proteins or carbohydrates. The probe is useful in various diagnostic and purification methods wherein hybridization of the oligonucleotide with a target nucleic acid is possible. In one instance, the probe can be used to capture a DNA strand which has been amplified using polymerase chain reaction techniques.

Nucleic acid probe, test kit and diagnostic and purification methods

Growing community of inventors

Rochester, NY, United States of America

John Bruce Findlay