Becton Dickinson and Company Limited

Dolores M Berger

William A Nussbaumer

Tobin J Hellyer

James G Nadeau

Robert W Rosenstein

Keith Edward Thornton

Andrew Kuhn

Sha-Sha Wang

Paul A Foxall

Klaus W Berndt

Paul Gerard Gladnick

Thomas L Fort

Gregory Tice

Paul Ernest Goldenbaum

Daretta A Yursis

Anne B Brown

Silvia A Bustos

Christine Ann Rostkowski

Sha Sha Wang

Karen Ann Dilly

Keith Thornton

Robert Rosenstein

A high-sensitivity, low-background immuno-amplification assay is provided, which offers a streamlined workflow suitable for high-throughput assays of clinically relevant samples, such as blood and other bodily fluids. The assay comprises the use of two proximity members that each comprise an analyte-specific binding component conjugated to an oligonucleotide. Binding an analyte brings the oligonucleotide moieties of the proximity members in sufficiently close contact that the oligonucleotides form an amplicon. The presence of the analyte then is detected through amplification of the amplicon and detection of the amplified nucleic acids. The sensitivity of the assay of the present invention is improved by preventing spurious or non-specific amplicon formation by proximity members that are not complexed with an analyte.

Immuno-amplification

The present invention relates to a method and composition for stabilizing clinical specimens (i.e., cells in biological samples) for transport and subsequent testing for diagnosis. The composition is specifically capable of maintaining nucleic acid in the cells intact for hybridization with oligonucleotide capture and detector probes.

Composition for providing long term stability to cells for diagnostic testing

Primers and probes derived from the HBV DNA polymerase gene which facilitate detection and/or quantification of all presently known genotypes of HBV. Disclosed sequences may be used in a variety of primer and probe constructs for detection of HBV nucleic acids.

Sequences and methods for detection of hepatitis B virus

A region of the,cryptic plasmid has been identified which is useful for performing amplification assays to determine specifically whether,is present in the sample being tested. Oligonucleotides useful for performing thermal Strand Displacement Assay (tSDA) reactions on this gene are disclosed. The disclosed oligonucleotides can be used in an assay which is specific for all strains of,and which does not show crossreactivity with the genomes of other microorganisms or with human DNA.

Assay for Chlamydia trachomatis by amplification and detection of Chlamydia trachomatis cryptic plasmid

A region of the Chiamydia trachomatis cryptic plasmid has been identified which is useful for performing amplification assays to determine specifically whether C. trachomiltis is present in the sample being tested. Oligonucleotides useful for performing thermal Strand Displacement Assay (tSDA) reactions on this gene are disclosed. The disclosed oligonucleotides can be used in an assay which is specific for all strains of C. trachomatis and which does not show crossreactivity with the genomes of other microorganisms or with human DNA.

Assay for Chlamydia trachomatis by amplification and detection of

Amplification primers and methods for complex-specific amplification of target sequences in the dnaJ genes of the Mycobacterium avium Complex (MAC) species are disclosed. The primer target binding sequences are useful for amplification of the dnaJ target in a variety of amplification reactions, with detection of the complex-specific target and, optionally, identification of the MAC species from which the target is derived. Primers and methods for multiplex amplification of dnaJ and a second target are also described.

Amplification and detection of mycobacterium avium complex species

Methods and apparatus for detecting biological activities in a specimen such as blood are disclosed. The present invention provides a system whereby the intensity of light with a wavelength within the range of about 600 nm to 800 nm introduced into a sample at a first location is measured as it reemerges at a second location. A significant change in the intensity of the reemerging light indicates the presence of biological activity. Preferably, the intensity is also measured at a fourth location, or alternatively, a second light source is disposed at a third location to permit comparative intensity data to be collected. These data are useful in partially identifying the types of microorganisms present in the sample. In preferred embodiments, light emitting diodes are used as the light sources and multiplexed to a plurality of samples. These signals are either detected using photodetectors and demultiplexed, or are collected using fiber-optic light guides and fed to a photomultiplier tube which generates an intensity signal. In either embodiment, the intensity signal is preferably amplified, digitized and stored in a computer.

Methods and apparatus for detecting biological activities in a specimen

A non-ionic lytic agent, preferably saponin for reducing the background carbon dioxide produced by blood cell metabolism is used in the testing of cultures for the presence of microorganisms. The hemolytic agent saponin is combined with a phospholipid, preferably L-.alpha.-Lecithin (phosphatidylcholine), to form mixed micelles which protect saponin from the effects of heat sterilization and high blood cholesterol levels, thus maintaining the lytic activity of saponin. The phospholipid/saponin mixed micelles are added to non-radiometric culture media vials such as Bactec.RTM. NR6A, NR7A and NR8A. The media vials are used in the Bactec.RTM. NR-660 and NR-730 instruments. However, the present invention may also be used in radiometric media such as Bactec.RTM. culture vials 6, 7 and 8 for reducing background carbon dioxide levels detected in C.sup.14 radiometric instruments such as the Bactec.RTM.-460, or in other instruments designed to detect bacteria in the presence of mammalian cells by monitoring the increase in CO.sub.2 produced by the metabolizing bacteria.

Growing community of inventors

Baltimore, MD, United States of America

Dolores M Berger