Abstract: HLA class I and class II locus specific DNA probes are prepared by cloning DNA segments obtained from the 3'-untranslated region and the 5' flanking region of isolated alleles. The probes are useful to determine the HLA alleles present in an individual.

Abstract: An improvement in the sensitivity of hybridization assays for detecting polynucleotide target sequences is provided by selective dehybridization of the bound portion of the probe, separation of the dehybridized portion from the support and the residual sample, and concentrating the separated probe before detecting its presence by means of its reporter group. The concentration step can be carried out by adsorption of the dehybridized probe onto a basic ion exchange material. A displacer sequence having greater binding affinity for the target polynucleotide sequence may be used to dehybridize the bound probe portion for subsequent concentration.

Abstract: A chemical agent is provided for significantly preventing or blocking non-specific staining or binding of an antibody specific for Terminal deoxynucleotidyl Transferase (TdT) during immunofluorescent or immunoperoxidase assay procedures. These procedures include both immunofluorescent and immunohistochemical staining of samples followed by flow cytometric and/or microscopic analysis, respectively. The invention is practiced by selective use of casein introduced into the assay procedures at an appropriate interval prior to analysis using a labelled or tagged monoclonal antibody specific to a TdT epitope. The casein utilized successfully was obtained from a large variety of sources and includes the use of a non-fat milk product.

Abstract: The invention concerns a probe for the detection of shigellae and entero-invasive E. coli, containing a nucleic acid sequence originating from the 140 Mdal virulence plasmid of the M 90 T strain of Shigella flexneri; having a maximum size of around 27 kb and inlcuding all or part of the 27 kb Bam HI fragment.This probe permits the in vitro diagnosis of syndromes of dysentery or diarrhea, of the Shigellosis type.

Abstract: A method and composition for use in preparing red blood cells for use as indicator cells to detect the presence of antigens and antibodies in biological fluids is the subject of the present invention. Freshly collected human or animal erythrocytes are washed six times with isotonic saline, and suspended to a hematocrit of four percent (volume/volume) with isotonic saline. An incomplete antibody, specific for the antigen or antibody to be detected, in a quantity less than that which would cause irreversible agglutination, is added to the erythrocyte solution. The mixture is incubated, washed, and suspended to a hematocrit of 3-5 (v/v) with isotonic saline. A second antibody, immuno specific for the first antibody and in an amount sufficiently great to immunologically bind substantially all of the antibody carrying red blood cells but not so great as to cause irreversible agglutination, is attached to the erythrocytes. This mixture is incubated, washed, and suspended to a hematocrit of 0.

Abstract: Recombinant human prealbumin and a method for producing such human prealbumin through recombinant DNA techniques including preparation and isolation of a cDNA sequence coding for human prealbumin, construction of a cloning vector and an expression vector containing said cDNA sequence, and expression of human prealbumin in a prokaryotic cell transformed by said expression vector are disclosed. The invention further relates to the use of human prealbumin cDNA in the diagnosis by hybridization methodologies of medical conditions with which variant forms of prealbumin are associated. Also disclosed is a method for diagnosing some Type I familial amyloid polyneuropathies by a restriction endonuclease assay with an enzyme which recognizes the nucleotide base sequence 5'-ATGCAT-3'.

Abstract: Novel compositions and methods are provided for detecting lupus nephritis in patients suffering from SLE. Competitive or non-competitive assays may be employed, where monoconal antibodies may be used to compete with antiserum for a nucleic acid ligand or may be used for the detection of antiidiotype antibodies as diagnostic of the presence or absence of lupus nephritis.

Abstract: This invention is directed to the use of specific polymethine dyes for staining fixed biopsy specimens of hematopoietic origin consisting of blood, bone marrow and lymph node cells for purposes of identifying and enumerating the stained cells when viewed under dark field illumination. The polymethine dyes, e.g. Astrazon Red Violet FRR, stain the lymphocyte subpopulations distinct colors capable of being viewed under dark field illumination for purposes of distinguishing one stained cell from another. This method of staining and the compositions thereof represents an advance in the art of lymphocyte subpopulation identification which was achieved heretofore primarily by using monoclonal antibody techniques.

Abstract: Chronic immune thrombocytopenic purpura is due to platelet destruction by circulating anti-platelet antibody. Although autoantibodies against the platelet glycoprotein IIb/IIIa complex and glycoprotein Ib have been demonstrated using various methods, practical assays for detection of platelet-associated or plasma autoantibodies have not been available. I studied 44 patients with chronic immune thrombocytopenic purpura where platelet-associated and plasma autoantibodies against the glycoprotein IIb/IIIa complex and glycoprotein Ib were measured using a newly developed immunobead assay and a previously reported microtiter well assay. Platelet-associated autoantibody was detected using the immunobead assay in nine of 11 patients (81.8%; seven with anti-GPIIb/IIIa, two with anti-GPIb). Plasma autoantibodies were noted in 28 of 44 patients (63.6%; 19 with anti-GPIIb/IIIa, seven with anti-GPIb and two with both).

Abstract: The invention offers an early detection method for atherosclerosis using genetic analysis to detect a polymorphisms shown to be correlated with this disease which are proximal to the apolipoprotein AI (apoAI) and aplipoprotein CIII (apoCIII) gene complex. All individuals with a 300 bp deletion 4 kb upstream of the apoAI gene are destined to experience severe atherosclerotic symptomologies. Individuals with a polymorphism 5.4 kb 5' of the apoAI gene or a PvuII polymorphism in the first intron of the apoCIII gene also seem to be at greater risk. A haplotype with MspI and XmnI/7.2 polymorphisms in this general region seem to be protected. Additional polymorphic sites in the DNA sequence associated with the apoAI/CIII gene complex provide a means for genetically fingerprinting individuals, and for identifying persons at risk with respect to disorders relating to lipid metabolism and transport.

Abstract: A diagnostic assay support matrix is prepared by coating a support matrix with a polymer resistant to nonspecific protein binding and immobilizing the desired biologically active agent on the polymer surface.

Abstract: A method for distinguishing fragments of DNA which contain single base mismatches from their perfectly paired homologues is disclosed. Single stranded regions within a duplex fragment are modified with carbodiimide, which reacts with unpaired guanine (G) and thymine (T) residues in DNA. Linear duplex DNA molecules do not react, while DNA molecules with single base mismatches react quantitatively with carbodiimide. Following reaction with carbodiimide, the DNA molecules are fractionated on high percentage polyacrylamide gels such that modified and unmodified fragments can be clearly distinguished. Application of this technique in order to located and purify DNA sequence differences responsible for phenotype variation and inherited disease is disclosed.

Abstract: Liquid loaded pads useful as wound and burn dressings are prepared from pellicles of microbially-produced cellulose obtained, for example, by culturing Acetobacter xylinum. A pellicle having a thickness from about 0.1 to 15 millimeters or greater is processed to replace the culture medium with water or other physiologically compatible liquid. The liquid-loaded pellicle is sterilized prior to its use as a dressing or in other medical applications.

Abstract: A method is disclosed for determining an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp) consisting of ligand and complementary receptor. The method comprises combining in an aqueous medium (1) said sample, (2) a first reagent comprising a non-dispersed surface to which is bound an sbp member that becomes bound to the surface in relation to the amount of analyte in the sample. The volume of the aqueous medium containing the above reagents is sufficiently large to allow complete immersion of the first reagent therein and determination of the analyte but not so large to result in substantial interference in the determination upon addition of a second reagent reactive with the conjugate of enzyme and sbp member and capable of generating a signal. The second reagent is present in a medium of sufficient volume to substantially increase the volume of the liquid medium and reduce interference in the determination.

Abstract: An immunoassay in which a thermally induced phase separation is used to effect the separation of specifically bound reactants from free reactants is disclosed. A first reactant is conjugated to a temperature-sensitive polymer to form a polymer/reactant conjugate, and a second reactant is conjugated to a reporter to form a reporter/reactant conjugate. The polymer/reactant, reporter/reactant, and biological fluid samples suspected of containing the analyte are admixed in solution at a temperature other than that at which the polymer will precipitate. Specific binding is allowed to occur, thereby forming a ternary complex. The salt concentration of the adjusted solution is then adjusted to a concentration sufficient to cause the complex to precipitate from the solution, the amount of reporter activity in the precipitated complex or in the solution measured and the presence and/or concentration of the analyte therefrom determined.

Abstract: The method of measuring circulating antigens or antibodies by using a ligand labeled specific antigen or ligand labeled specific antibody chemically attached to a soluble matrix or backbone, a differently labeled anti-antigen or anti-antibody and a solid phase anti-ligand directed at the ligand attached to the specific antigen or specific antibody. This is achieved by either one or two analytical schemes:(a) Reacting a patient sample with a ligand labeled specific antigen or a ligand labeled specific antibody in the liquid phase in the presence of a differently labeled specific anti-antigen or labeled specific anti-antibody. This immunological complex is reacted with an immobilized anti-ligand on a solid support which is directed against the ligand attached to the specific antigen or antibody through the liquid matrix. Subsequently the solid phase is washed and checked for the label on the anti-antigen or anti-antibody which is directly proportional to the concentration of specific antigen or antibody.

Abstract: A heterogeneous specific binding assay method for determining the amount of a suspected analyte in an aqueous test medium wherein a reaction mixture is formed by combining the test medium with assay reagents including a labeled reagent, an immobilizable component, and a binding substance which causes the immobilizable component to precipitate. Free and bound species of the labeled reagent are formed as a function of the amount of the analyte in the test medium. One of the free and bound species of the labeled reagent is immobilized by binding of the immobilizable component with the binding substance. The immobilized labeled reagent is seperated from labeled reagent which has not been immobilized, and the amount of label in the labeled reagent in one of the separated fractions is determined and related to the amount of analyte in the test medium.

Abstract: Polymorphisms in genes related to lipid metabolism, specifically apolipoproteins B, CII, E, and apoAIV, have been identified. Presence or absence of these polymorphisms in particular individuals may be correlated with propensity to show symptoms of atherosclerosis. Also thus correlated are two insertion polymorphisms 5' of the insulin gene.

Abstract: Nucleic acid hybridization probes are provided which have sequences complementary to sequences of segments in bovine male-specific DNA and are suitable for sexing bovine embryos at the time of embryo transfer with virtually 100% accuracy.

Abstract: Fragments of human Factor VIII:C which bind to antibody inhibitors of Factor VIII in patients afflicted with such inhibitors are disclosed. The method of treating Factor VIII inhibitors, by administering one or more of these fragments, is also disclosed.