Abstract: Strand Displacement Amplification methods (thermophilic SDA) which can be performed over a broad temperature range (37.degree. C. to 70.degree. C.). The preferred temperature range for thermophilic SDA is 50.degree. C. to 70.degree. C. It has been found that certain thermophilic restriction endonucleases are capable of nicking the hemimodified restriction endonuclease recognition/cleavage site as required by SDA and dissociating from the site. It has further been found that certain thermophilic polymerases are capable of extending from the nick while displacing the downstream strand. Thermophilic SDA, because of reaction temperatures higher than previously possible with conventional SDA enzyme systems, has improved specificity and efficiency, reduced nonspecific background amplification, and potentially improved yields of amplification products.

Abstract: The invention is directed to a method of detecting infection of a blood sample by various strains of bovine diarrhoea virus (BVD) including a first test wherein anti-bovine diarrhoea virus antibodies are detected by means of a recombinant antigen comprising the BVD p80 protein expressed by a eukaryotic host and a second test wherein viral particles are detected by means of antibodies directed against the BVD p80 viral protein. The method is particularly useful in detecting pathogenic conditions such as persistent viremias and acute infections caused by BVD viruses, and is particularly advantageous because it is an extremely sensitive and efficient test and because it is capable of detecting infections caused by any one of a wide variety of BVD strains. The recombinant antigen employed in the method is preferably encoded by the nucleotide sequence listed as SEQ ID No: 1.

Abstract: Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

Abstract: Methods and reagents for determining an individual's genotype at the Glycophorin A locus with respect to a set of five alleles using sequence-specific oligonucleotide probes enable one to type samples from a variety of sources, including samples comprising RNA or cDNA templates, and can be applied to nucleic acids in which a target region spanning the polymorphism has been amplified. This typing facilitates typing tissue for determining individual identity and has application in the field of forensic science.

Abstract: A method of diagnosing cat scratch disease and a method of diagnosing bacillary angiomatosis in a subject by detecting the presence of Rochalimaea henselae or an immunogenically specific determinant thereof in the subject is provided. Also provided is a vaccine comprising an immunogenic amount of a nonpathogenic Rochalimaea henselae or an immunogenically specific determinant thereof and a pharmaceutically acceptable carrier. A method of diagnosing Rochalimaea quintana infection in a subject by detecting the presence of a nucleic acid specific to Rochalimaea quintana in a sample from the subject is provided. A purified heat shock protein of Rochalimaea is provided.

Abstract: An isolated nucleic acid molecule comprising the agfA gene of Salmonella. Methods and compositions suitable for diagnostic tests utilizing the isolated gene, and protein therefrom, to give highly specific diagnostic assays to Salmonella, and/or enteropathogenic bacteria of the family Enterobacteriaceae.

Abstract: Thermophilic Strand Displacement Amplification (tSDA) for amplification of nucleic acid target sequences in situ in cells in suspension, on slides or in tissues is described. Excellent specimen morphology is preserved, and either DNA targets, RNA targets, or both may be selectively amplified. In situ amplification by tSDA is compatible with immunochemical techniques, so that both amplification of target sequences and immunological staining can be performed on the same specimen.

Abstract: The present invention is directed to methods and reagents for the specific detection and presumptive identification of various bacteria associated with waterborne infectious disease. In particular, this invention relates to methods and reagents for the specific detection and identification of Salmonella and Shigella in environmental samples such as water and sewage.

Abstract: The present invention provides a nucleic acid encoding human Astrovirus serotype 2, or a unique fragment thereof. The sequence, a genomic RNA of human astrovirus serotype 2 contains 6,797 nucleotides, and is organized into three open reading frames. Also provided are purified antigenic polypeptide fragments encoded by the nucleic acid encoding human Astrovirus serotype 2, or unique portions thereof. The present invention also provides a monoclonal antibody specific for human astrovirus serotype 2 and isolated nucleic acids capable of selectively hybridizing with the nucleic acid of serotype 2, including methods for detecting the presence of serotype 2 utilizing these products.

Abstract: Amplification and/or expression of nucleic acids is carried out in a medium immobilized by using an organic and/or inorganic solid matrix penetrating the medium and having a porous, fibrous, reticulated, coiled, capillary, lamellar or folded texture and which includes the components of a cell-free enzyme system of exponential amplification of nucleic acids and/or components of a cell-free enzyme system of nucleic acid expression. In this medium, the progeny of each molecule (clone) and the expression products remain in the same zone of the reaction volume where the matrix molecule was initially located. The method permits cloning of nucleic acids in vitro as well as detection of solitary nuleic acid molecules in the sample studied.

Abstract: The dependence of ionizing radiation-induced GADD45 mRNA expression on the presence of functional p53 in mammalian cells is disclosed. First and second oligonucleotide sequences are provided which can form a double-stranded oligomer capable of binding to functional p53 protein. The present invention demonstrates that the dependence of ionizing radiation-induced GADD45 mRNA expression on the presence of functional p53 and the binding of functional p53 to a double-stranded oligomer binding sequence can serve as the basis for methods for determining the presence of functional p53 in mammalian cell lines and tumors.

Abstract: Mutant ribozymes are screened by culturing cells whose survival is dependant upon cleavage of RNA by a ribozyme, which cleavage causes the cells to survive in the presence of an agent which otherwise would kill the cells, and selecting cells which survive.

Abstract: The present invention is directed to compositions comprising an oligonucleotide linked to a detectably labeled marker component comprising a fluorophore moiety which comprises a substantially planar, multidentate macrocyclic ligand coordinated to a central atom capable of coordinating with two axial ligands and two polyoxyhydrocarbyl moieties which are attached as axial ligands to the central atom. The present invention is also directed to nucleic acid hybridization and amplification methods employing such compositions.

Abstract: A human gene has been discovered which is genetically altered in human tumor cells. The genetic alteration is gene amplification and leads to a corresponding increase in gene products. Detecting that the gene, designated hMDM2, has become amplified or detecting increased expression of gene products is diagnostic of tumorigenesis. Human MDM2 protein binds to human p53 and allows the cell to escape from p53-regulated growth.

Abstract: Polynucleotide sequences encoding enzymes that possess .alpha.2-3 neuraminidase activity are provided. Of particular interest are polynucleotide sequences encoding an enzyme having .alpha.2-3 neuraminidase activity and naturally produced by the bacteria Streptococcus pneumoniae. Recombinant DNA expression of enzymes possessing .alpha.2-3 specific neuraminidase activity is also described, including methods, recombinant host cells and a genetic construction. The invention also provides a purified enzyme having .alpha.2-3 neuraminidase activity from Streptococcus pneumoniae, wherein the enzyme is isolated from S. pneumoniea cultures. Another aspect of this invention is to provide methods of isolating genes encoding enzymes having neuraminidase activity, particularly .alpha.2-3 neuraminidase activity. The gene isolation methods of the invention comprise the step of labeling a hybridization probe derived from the neuraminidase coding portion of plasmid pND-1.

Abstract: The ability to rapidly detect wild polioviruses in clinical specimens is a major concern for the world-wide eradication of polioviruses. This report describes a method of detecting polioviruses of all three serotypes from viral isolates of clinical specimens using a pair of degenerate PCR primers. This primer set, which uses deoxyinosine residues to compensate for third position mismatches, recognizes nucleotide sequences near the receptor binding site of polioviruses. These sequences are unique to polioviruses and are absolutely conserved at the amino acid level. As a result, these PCR primers do not recognize nonpoliovirus enteroviruses. All poliovirus serotypes (40 poliovaccine related genotypes and 120 wild poliovirus genotypes from around the world) tested positive. All 14 prototype strains of nonpoliovirus enteroviruses tested negative. This "pan-poliovirus" degenerate PCR primer set will be useful in rapidly diagnosing poliovirus infections from world-wide clinical specimens.

Abstract: The invention provides a method, compositions, and kits useful for detecting mycobacteria in a sample. The method includes contacting the sample with a formaldehyde solution, an organic solvent, and a protein-degrading agent prior to hybridizing a mycobacteria-specific nucleic acid probe to the sample. The invention has particular utility in detection and susceptibility screening of human-disease causing mycobacteria such as Mycobacterium tuberculosis.

Abstract: 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.

Abstract: An improved method for detecting and isolating differentially expressed mRNAs which comprises using first oligonucleotide primers for reverse transcription of mRNAs and both the first oligonucleotide primers and second oligonucleotide primers for amplification of the resultant cDNAs. The improvement of this method comprises providing first and second oligonucleotide primers with a length of at least 21 oligonucleotides. The method further comprises using a two-step PCR amplification, wherein non-stringent conditions are used for the first 1 to 4 cycles, and stringent conditions are used for the next 16 to 22 cycles. This highly reproducible method will permit the preparation of comprehensive catalogs of gene expression for any given cell type.

Abstract: There is provided by this invention methods of detecting genetic deletions and mutations associated with at least one condition selected from the group consisting of DiGeorge syndrome, Velocardiofacial syndrome, CHARGE association, conotruncal cardiac defect, and cleft palate in a human patient comprising the steps of providing a DNA containing test sample from said human patient; identifying whether there are less than two functional copies of the DiGeorge syndrome critical region loci, whereby said identification of less than two copies of the DiGeorge syndrome critical region loci is indicative of a likelihood that said person has at least one of DiGeorge syndrome, Velocardiofacial syndrome, CHARGE association, conotruncal cardiac defect, and cleft palate. Probes and primers useful in the invention are also provided as are diagnostic kits.