Abstract: Recombinant human neurokinin-1 receptor (human NKIR) is disclosed which has been isolated by polymerase chain reaction techniques. Also disclosed is the complete sequence of human NKIR cDNA, expression systems containing said isolated cDNA, including a CHO (Chinese Hamster Ovarian Cell Line) stable expression systems, and an assay using the CHO eexpression system. NKIR, also known as substance P receptor, can be used in an assay to identify and evaluate entities that bind substance P receptor. The assay can also be used in conjunction with diagnosis and therapy to determine the body fluid concnetration of substance P in arthritis patients.

Abstract: A binding assay process for an analyte, using a capture binding agent with binding sites specific for the analyte and a developing binding material capable of binding with the bound analyte or with the binding sites on the capture binding agent either occupied by the bound analyte or the remaining unoccupied binding sites, employs the capture binding agent in an amount such that only an insignificant fraction of the sample analyte becomes bound to the capture binding agent, which is preferably provided at high surface density on microspots. A label is used in relation to the developing binding material and is provided by microspheres which are less than 5 .mu.m and carry a marker preferably fluorescent dye molecules. To determine the concentration of sample analyte, the signal strength, which represents the fractional occupancy of the binding sites on the capture binding agent by the analyte, is compared with a dose-response curve computed from standard samples.

Abstract: Enzymatic synthesis of a portion of an oligonucleotide is performed by a cycle of synthetic steps: (a) combining an oligonucleotide primer and a blocked nucleotide in a reaction mixture in the presence of a chain extending enzyme, such that a primer-blocked nucleotide product is formed, wherein the blocked nucleotide substrate comprises (i) a nucleotide to be added to form part of the defined sequence and (ii) a blocking group attached to the nucleotide effective to prevent the addition of more than one blocked nucleotide to the primer; and (b) removing the blocking group from the 3' end of the primer-blocked nucleotide product to form a primer-nucleotide product. Phosphate is generated in at least one synthetic step. A precipitate is formed in the cycle comprising phosphate and at least one precipitation cation. The precipitation of phosphate reduces its unfavorable effect on the method. Preferably, cycles of the method are repeated without intermediate purification of primer-nucleotide product or precursor.

Abstract: Oligonucleotides and methods for the amplification and specific detection of Chlamydia trachomatis. The invention relates to amplification oligonucleotides capable of amplifying Chlamydia trachomatis nucleotide sequences and to probes and helper oligonucleotides for the specific detection of Chlamydia trachomatis nucleic acids. The invention also relates to methods for using the oligonucleotides of the present invention and specific combinations and kits useful for the detection of Chlamydia trachomatis.

Abstract: Compositions and methods for covalently immobilizing an oligonucleotide onto a polymer-coated bead or similar structure are described. Specifically, the polymer-coated bead or similar structure possesses a large number of activatable moieties, preferably primary and secondary amines. An oligonucleotide is activated with a monofunctional or multifunctional reagent, preferably the homotrifunctional reagent cyanuric chloride. The resultant covalently immobilized oligonucleotides on the beads or similar structures can serve as nucleic acid probes on solid supports, and hybridization assays can be conducted wherein specific target nucleic acids are detected in complex biological samples.

Abstract: A method for determining head and neck squamous cell carcinomas, bladder tumors and prostate carcinomas is described. The method involves assaying for expression of the gene coding for tumor rejection antigen precursor MAGE-3, or its expression product. Various assays, and kits useful for these assays, are described.

Abstract: Methods and reagents are provided for the amplification of DNA sequences longer than 10 kilobases by the polymerase chain reaction (PCR). The methods use compositions consisting of a primary thermostable DNA polymerase from Thermus thermophilus combined with a lesser amount of a secondary thermostable DNA polymerase possessing a 3'-to-5' exonuclease activity from Thermococcus litoralis, Pyrococcus species GB-D or Thermotoga maritima. The DNA polymerase compositions, when used with the disclosed reaction buffer, enable amplifications of DNA sequences up to at least 42.2 kilobases in length.

Abstract: A method for screening for bladder cancer by identifying expression of one or more of MAGE-1, MAGE-2, MAGE-3 and MAGE-4 is the disclosed invention. Expression can be determined by a number of methods, including nucleotide amplification assays.

Abstract: The invention provides a rapid process for lysing Mycobacteria. In one embodiment is provided a process for lysing Mycobacteria which comprises exposing the bacteria to a lysis effective amount of heat. A particularly effective method for providing the necessary heat is in the form of forced hot air such as in a forced hot air oven. The process of the invention is particularly advantageous since only one step is involved, it is expedient compared to prior methods, and little instrumentation is necessary. By practicing the present invention it is possible to lyse Mycobacteria with minimal effort. In addition, practicing the invention results in liberating cellular components including deoxyribonucleic acid (DNA) from Mycobacteria. Not only is DNA liberated, but the DNA is suited for subsequent analysis by way of probe hybridization, restriction enzyme analysis, and the like.

Abstract: Oligonucleotides and methods for the amplification and specific detection of Chlamydia trachomatis. The invention relates to amplification oligonucleotides capable of amplifying Chlamydia trachomatis nucleotide sequences and to probes and helper oligonucleotides for the specific detection of Chlamydia trachomatis nucleic acids. The invention also relates to methods for using the oligonucleotides of the present invention and specific combinations and kits useful for the detection of Chlamydia trachomatis.

Abstract: The present invention provides methods and reagents for the detection and identification of four causative agents of genital ulcerations, herpes simplex virus (HSV) types 1 and 2, Treponema pallidum, and Haemophilus ducreyi. The methods use sequence-specific primers which enable the simultaneous polymeric chain reaction amplification of genomic nucleic acid sequences from HSV types 1 and 2, T. pallidum, and H. ducreyi. Following amplification, sequence-specific oligonucleotide probes are used to detect and distinguish HSV type 1 and 2, T. pallidum, and H. ducreyi nucleic acid.

Abstract: The method of the present invention relates to a rapid procedure for detecting DNA in a cell, while preserving the morphology of the nucleus for analysis. The method comprises depositing a cell onto a polymeric membrane filter wherein the DNA contained in the cell is retained on the polymeric membrane filter and is available for binding with a fluorescently labeled nucleic acid probe, incubating the polymeric membrane filter with the labeled nulceic acid probe, and detecting the labeled nulceic acid probe wherein detection of the nulceic acid probe is indicative of the presence of the DNA. There are no separate permeabilization steps needed to permit the probes to enter the cell and hybridize to the DNA. The method is simple and quick and is applicable to the sample volumes found in clinical laboratories.

Abstract: The present invention is directed to methods for controlling the light emission of an oligonucleotide labeled with a light-emitting label that are useful in nucleic acid detection assays. A reaction that results in the cleavage of single-stranded oligonucleotide probes labeled with a light-emitting label is carried out in the presence of a DNA binding compound that interacts with the label to modify the light emission of the label. The methods utilize the change in light emission of the labeled probe that results from degradation of the probe. The methods are applicable in general to assays that utilize a reaction that results in cleavage of oligonucleotide probes, and, in particular, to homogeneous amplification/detection assays wherein hybridized probe is cleaved concomitant with palmer extension. A homogeneous amplification/detection assay is provided which allows the simultaneous detection of the accumulation of amplified target and the sequence-specific detection of the target sequence.

Abstract: The present invention encompasses a method of synthesis of conjugates of photoproteins that retain all or a substantial portion of the luminescent activity of underivatized photoprotein. According to the present invention photoproteins may be conjugated with a variety of binding including streptavidin/avidin, glyco-proteins, lectins, hormones, antigens, drugs, antibodies and antigen binding fragments thereof, or any other selectively bindable reagent by chemical crosslinking means. The present invention also encompasses conjugates produced by this method, and methods of use of such conjugates.