Abstract: A method is described for diagnosing individuals as having hypertrophic cardiomyopathy, e.g. familial or sporadic hypertrophic cardiomyopathy. The method provides a useful diagnostic tool which becomes particularly important when testing asymptomatic individuals suspected of having the disease. Symptomatic individuals have a much better chance of being diagnosed properly by a physician. Asymptomatic individuals from families having a history of familial hypertrophic cardiomyopathy may be selectively screened using the method of this invention allowing for a diagnosis prior to the appearance of any symptoms. Individuals having the mutation responsible for the disease may be counseled to take steps which hopefully would prolong their life, i.e. avoid rigorous exercise. The methodology used in the above method also has broad applicability and may be used to detect other disease-associated mutations in DNA obtained from subjects being tested for other disease-associated mutations.

Abstract: A primer directed DNA amplification method to isolate efficiently chromosome-specific repeated DNA wherein degenerate oligonucleotide primers are used is disclosed. The probes produced are a heterogeneous mixture that can be used with blocking DNA as a chromosome-specific staining reagent, and/or the elements of the mixture can be screened for high specificity, size and/or high degree of repetition among other parameters. The degenerate primers are sets of primers that vary in sequence but are substantially complementary to highly repeated nucleic acid sequences, preferably clustered within the template DNA, for example, pericentromeric alpha satellite repeat sequences. The template DNA is preferably chromosome-specific. Exemplary primers ard probes are disclosed. The probes of this invention can be used to determine the number of chromosomes of a specific type in metaphase spreads, in germ line and/or somatic cell interphase nuclei, micronuclei and/or in tissue sections.

Abstract: Hepatitis C virus oligonucleotides and a system to determine hepatitis C virus genotypes with polymerase chain reaction utilizing those oligonucleotides as primers.

Abstract: A method of characterizing the chromosomes in a sample of cells by fixing the cell sample on a substrate, contacting the cell sample with a nucleic acid probe having a detectable label under conditions that allow the probe to hybridize preferentially to a chromosome in the cells to form a hybridized complex, optically detecting each labeled complex in the sample, defining a predetermined number of neighboring labeled complexes as a group, generating a distance parameter based on the distance between the position of a group and the position of the next neighboring labeled complex, and comparing the distance parameter for each group to a standard distance value to characterize the chromosomes in the cells of the sample.

Abstract: The subject invention relates to A set of DNA primers which, when utilized in conjunction with the polymerase chain reaction (PCR) assay, can amplify and speciate DNA from four medically important Candida species. Furthermore, the PCR amplified products, generated by the primers, can also be used to create species specific probes which can also detect and confirm the four species of Candida. Thus, the present invention allows for early diagnosis and treatment of an infection.

Abstract: The present invention relates to species-specific DNA probes specific for Vibrio vulnificus and Vibrio cholerae. The DNA probes of the present invention specifically detects Vibrio vulnificus or Vibrio cholerae in a mixed bacterial sample based on unique ribosomal RNA nucleotide sequences. When the DNA probes of the present invention are tagged with a labeled molecule such as a fluorescent label, it affords direct and immediate visualization of individual bacterial cells, and a rapid method of detection of bacterial infection in humans and shellfish without culturing.

Abstract: The present invention provides single-stranded circular oligonucleotides each with at least one parallel binding (P) domain and at least one corresponding anti-parallel binding (AP) domain separated from each other by loop domains. When more than one P or AP domain is included in a circular oligonucleotide of the present invention, the additional P or AP domains can constitute loop domains for a pair of corresponding P and AP domains, and vice versa. Each P and AP domain has sufficient complementarity to bind to one strand of a defined nucleic acid target wherein the P domain binds in a parallel manner to the target and the corresponding AP domain binds in an anti-parallel manner to the target. Moreover, the present single-stranded circular oligonucleotides can bind to both single-stranded and double-stranded target nucleic acids. The present invention also provides methods of making and using these oligonucleotides as well as kits and pharmaceutical compositions containing these oligonucleotides.

Abstract: This invention relates to a method of cloning DNA produced by primer extension including PCR amplified, reverse transcriptase-generated or primer extended synthetic DNA. Specifically, it relates to a method in which alkane diol residue containing oligonucleotide primers are incorporated into DNA by primer extension followed by direct cloning of the target DNA. Following transformation, the host excises the alkane diol residue with its endogenous DNA repair machinery.

Abstract: Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequence defined in the Sequence Listing by SEQ ID NO:5. This is the C. albicans ITS2 sequence and includes a nucleic acid comprising a nucleotide sequence that is specific for Candida albicans. Further examples of an isolated double stranded nucleic acid of the present invention consist essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID NOs:6-9. These are the ITS2 sequences for C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium polyanetholesulfonate, and sodium ethylene diamine.RTM. tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.

Abstract: Method of diagnosing arterial chlamydial granuloma by detecting in a biological sample both a first marker associated with Chlamydia pneumoniae and a second marker associated with arterial granuloma. Therapeutic composition for treating arterial chlamydial granulomatous disease, including an anti-Chlamydia pneumoniae agent and a granuloma inhibitor.

Abstract: Non-primate or primate cells are provided comprising a functional human transporter for neurotransmitter uptake. The cells allow for dissection of the mechanism of neurotransmitter transport, as well as screening for agonists and antagonists of the neurotransmitter with respect to its uptake. Methods are provided for producing such cells. Specifically, the cells are transformed with human DNA comprising the gene encoding for the neurotransmitter transporter, whereby this protein(s) is expressed and incorporated into the plasma membrane and is capable of functioning to transfer the neurotransmitter from the extracellular space to intracellular domains. The physiological, kinetic and pharmacological characteristics of transport in these cells conform to known characteristics of high-affinity neurotransmitter transport in the CNS.

Abstract: The use of specific primers and a probe in a reverse transcriptase-polymerase chain reaction provides a method for specifically identifying bovine respiratory syncytial virus in cattle.

Abstract: Stop solutions including a compound selected from the group consisting of acetamide, propionamide, butyramide and N-methylacetamide, and methods and kits for their use, e.g., in DNA sequencing.

Abstract: A full length cDNA clone from a normal human mammary epithelial cell (strain 184) encoding a 25 kDa protein, HME1, was isolated. Expression of HME1 RNA appears to be limited to epithelial cells. The HME1 sequence shares strong sequence homology with bovine 14-3-3 protein, which is an activator of tyrosine and tryptophan hydroxylase, but the tissue distribution of HME1 differs from that of 14-3-3. Compared with normal mammary epithelial cells, expression of HME1 RNA is dramatically low in cells derived from human mammary carcinoma and in normal mammary epithelial cells transformed by oncogenes. HME1 therefore appears to be a cellular differentiation marker that is down-regulated during neoplastic transformation.

Abstract: Nucleic acid hybridization probes having at least one nucleic acid strand which has at least two separate target specific regions that hybridize to a target nucleic acid sequence, and at least two distinct arm regions that do not hybridize with the target nucleic acid but possess complementary regions that are capable of hybridizing with one another. These regions are designed such that, under appropriate hybridization conditions, the complementary arm regions will not hybridize to one another in the absence of the target nucleic acid; but, in the presence of target nucleic acid the target-specific regions of the probe will anneal to the target nucleic acid, and the complementary arm regions will anneal to one another, thereby forming a branched nucleic acid structure.

Abstract: Disclosed are methods and compositions for the preparation of steroid 5.alpha.-reductases by recombinant means, as well as for the use of these enzymes in screening assays for the identification of compounds which have the ability to inhibit or otherwise alter the enzymatic function of these enzymes. Biochemical and pharmacological evidence is presented to demonstrate the existence of more than one human steroid 5.alpha.-reductase. The DNA sequence encoding steroid 5.alpha.-reductase 2, the major active isozyme of human genital tissue, is disclosed herein, in addition to methods and compositions for its preparation and pharmacological analysis. The sequences disclosed herein may be used directly in the preparation of genetic constructs, or may be employed in the preparation of hybridization probes for the selection of enzyme-encoding sequences from other sources.

Abstract: A method of cleaving a target nucleic acid molecule is disclosed. A cleavage structure is formed that comprises the target nucleic acid and a pilot nucleic acid. A first region of the target nucleic acid is annealed to the pilot nucleic acid to form a duplex structure. A second region of the target nucleic acid contiguous to the duplex is not annealed to the pilot nucleic acid, thus forming a junction site between the duplex region and the non-annealed region. The cleavage structure is exposed to a cleavage agent capable of preferentially cleaving the cleavage structure at a target site in a manner independent of the sequence of the cleavage structure. The cleavage structure and the cleavage agent are incubated under conditions wherein cleavage can occur.

Abstract: 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.

Abstract: Methods for multiplex amplification of target nucleic acid sequences using a single pair of primers. Defined sequences are appended to the ends of multiple target sequences as part of the amplification reaction so that no steps in addition to amplification are required. The target sequences with the appended defined sequences need not be isolated prior to amplification. In one embodiment for coamplification of two target sequences, a sequence corresponding to a terminal segment of the first target sequence is appended to one end of the second target sequence and a sequence corresponding to a terminal segment of the second target sequence is appended to one end of the first target sequence. Amplification of the two targets then requires only a single pair of primers. Alternatively, a single defined sequence may be appended to the 5' and 3' ends of any number of selected targets. All such modified target sequences may then be amplified using a single pair of primers which hybridize to the defined end-sequences.

Abstract: The present invention is directed toward the human D.sub.4 dopamine receptor. The nucleotide sequence of the gene corresponding to this receptor is provided by the invention. The invention also includes a recombinant eukaryotic expression vector capable of expressing the human D.sub.4 dopamine receptor in cultures of transformed eukaryotic cells and such cultures of transformed eukaryotic cells which synthesize the human D.sub.4 dopamine receptor.