Abstract: This invention provides a measuring method for the quantitative flow injection analysis of metal elements contained in body fluids comprising introducing the protein liberated by the reaction of body fluid sample and protein-precipitating reagent into a separating membrane for preventing the passage of protein to separate and remove the liberated protein, and then introducing the reacted solution into quantitative analysis means for determining and measuring the concentration of target metal contained in the body fluid.

Abstract: This invention relates to a novel reagent and a method of using the reagent in an immunoassay to detect antigens, particularly antigens immunocomplexed with their corresponding or cross-reacting antibodies. In particular, this reagent and method increase the detection of human immunodeficiency virus HIV-1 p24 core antigen.

Abstract: Disclosed is a method for the direct determination of the free fraction of analytes present in biological fluids in a free form and in a form bound to one or more endogenous ligands (said free and bound forms being in equilibrium with one another). This method provides for a (preferably substantially simultaneous) use:a first ligand L1 capable of sequestering an analyte quantity proportionate to the free-analyte concentration present in a biological fluid and to subsequently release it, after removal from the biological fluid of the specific endogenous ligand, as a result of the addition of an appropriate selective dissociating agent; a second ligand capable of binding both the previously released analyte and a labelled version of the analyte, even in the presence of the dissociating agent; a selective dissociating agent; and a quantity of labelled analyte.

Abstract: Methods are disclosed for inactivating interfering binding proteins in a immunoassay for a member of a specific binding pair (sbp). The method comprises including in an assay medium containing a sample suspected of containing an sbp member and an interfering binding protein an effective amount of a water soluble compound having two substituted or unsubstituted phenyl groups linked to a common atom. When the sbp member or its sbp partner has two phenyl groups linked to a common atom, the compound has a number of groups other than hydrogen attached to the phenyl groups and the atom that differs by at least two from the number of such groups on the sbp member. When the sbp member or its sbp partner has two phenyl groups linked to a common atom and the binding protein is not an antibody, the compound has only one group other than hydrogen attached to a phenyl group or the common atom.

Abstract: The present invention relates to the measurement of estradiol using competitive immunoassay methods. The inventors unexpectedly discovered that estrone and its derivatives conjugated to a label is a particularly effective tracer when used in conjunction with estradiol specific antibodies to determine estradiol levels in fluid samples. The present invention also utilizes 5.alpha.-dihydrotestosterone to enhance the assay performance.

Abstract: The present invention provides an improved method for staining slides using immunochemical reagents. The method comprises the following steps. The assay region of a slide (the region containing the tissue section) is washed with an improved rinsing solution comprising water and a detergent. An evaporation inhibitor liquid is applied to the slide to cover the assay region. For antigens requiring unmasking, the tissue section is combined with an improved, stabilized proteolytic enzyme solution. The slide is rinsed, and the evaporation inhibitor liquid is reapplied to the slide. A primary antibody in an improved diluent containing globulins from the same species as a second antibody is combined with the tissue section for a time sufficient for substantially complete antibody binding. The slide is rinsed, and the evaporation inhibitor liquid is reapplied. A labeled second antibody in the improved diluent is combined with the tissue section for a time sufficient for substantially complete antibody binding.

Abstract: A system for determining the concentration of fructosamine in sera which consists of a first reagent in which a tetrazolium salt which reduces all reactive substances in sera including fructosamine and a second reagent which is responsive to all reactive substance in sera other than fructosamine. The difference in color change as between the two allows the determination of concentration of fructosamine.

Abstract: The invention concerns a method for detecting an analyte in a blood sample or a derived fraction thereof. In this method one uses the binding affinity between the analyte and its specific binding agent. Moreover, a reaction component containing a peroxidase label is added to the sample and specific binding agent to form a reaction mixture. The incubation of the reaction mixture, containing the analyte and its specific binding agent, with the peroxidase-labeled reaction component has to be carried out in the presence of a quaternary ammonium salt or ionic surfactant.

Abstract: A method and kit for the isolation and quantitation of glycated hemoglobin and other glycated proteins, specifically albumin, from a single sample of whole blood. Glycated hemoglobin is calculated from an eluate resulting from the contact of the whole blood hemolysate with a boronated resin. Glycated albumin and other plasma proteins are calculated using immunoturbidimetry, resulting from reaction between a buffered antibody solution and the albumin or other protein in the previous eluate.

Abstract: In order to determine an analyte by a heterogeneous immunological test using a first partner of a binding pair which is immobilized on a solid phase in which a sample solution containing the analyte is incubated in the presence of the solid phase with either(1) a first conjugate consisting of (a) the second partner of the above-mentioned binding pair and (b) a first substance capable of specific immunological binding to the analyte or(2) a first conjugate consisting of (a) the second partner of the above-mentioned binding pair and (b) a first substance capable of specific immunological binding to the analyte as well as with a second labelled substance capable of specific immunological binding to the analyte,afterwards the phases are separated, in case (1) the liquid phase is replaced by a further solution containing a second labelled substance capable of binding specifically to the analyte and it is again incubated and subsequently separated, and in both cases the label on the solid phase or in the liquid pha

Abstract: The present invention provides ligand/anti-ligand assays for detecting and/or measuring adherent proteins, including lipophilic serum or plasma proteins, such as serum amyloid A (SAA) and apolipoprotein A1 (apoA1); cytokines such as IL-1 beta, IL-6, and TNF alpha; pentraxins, such as CRP; and globular serum or plasma proteins such as albumin.

Abstract: Use of a lithium salt, particularly lithium thiocyanate, as an agent for lysing red blood cells and denaturing hemoglobin in the performance of an immunoassay to detect the relative amount of a hemoglobin derivative in a blood sample. The method is particularly useful in the determination of hemoglobin Alc. The method provides a rapid means for releasing hemoglobin from red blood cells by lysis and for exposing the epitope that is characteristic of the Alc form. Low concentrations of the lithium salt provide rapid release and denaturation of hemoglobin without the need for dilution prior to the performance of the immunoassay.

Abstract: Physiological samples are typed for HLA-B27 by contacting the sample with an antibody which binds to HLA-B7 and is not cross-reactive between HLA-B7 and B27, separating the sample from any complexes which are formed, and then testing the sample for the presence of HLA-B27 with an antibody which is cross-reactive for HLA-B7 and B27 in a STAT test. Particularly, an enzyme conjugate is used which binds to a membrane allowing for the detection of any HLA-B27 bound to the membrane.

Abstract: A kit for inactivating interfering binding proteins in a immunoassay for a member of a specific binding pair (sbp). The method comprises including in an assay medium containing a sample suspected of containing an sbp member and an interfering binding protein an effective amount of a water soluble compound having two substituted or unsubstituted phenyl groups linked to a common atom. When the sbp member or its sbp partner has two phenyl groups linked to a common atom, the compound has a number of-groups other than hydrogen attached to the phenyl groups and the atom that differs by at least two from the number of such groups on the sbp member. When the sbp member or its sbp partner has two phenyl groups linked to a common atom and the binding protein is not an antibody, the compound has only one group other than hydrogen attached to a phenyl group or the common atom.

Abstract: Processes for preparing stable natural matrices for prostrate specific antigen (PSA) are disclosed. Biological carrier fluids for PSA obtained from a suitable mammal are modified to inhibit the activity of components of the biological fluids destabilizing to PSA. The stable natural matrices, prepared in accordance with the present invention, are useful in the measurement of PSA in a sample by means of an immunoassay.

Abstract: A competitive immunoassay to measure Lp(a) levels semiquantitatively and identify individuals with elevated plasma Lp(a) protein levels (greater than 7 mg/dl) who have an increase risk for coronary artery disease. The assay is useful for monitoring the effectiveness of Lp(a)-lowering drugs. The assay provides a quick, reliable, easy to use and inexpensive method to measure plasma Lp(a) level with serum, plasma or whole blood.

Abstract: The present invention involves a method for detecting bladder cancer in a subject. The method preferably comprises first collecting a urine sample from the subject. The presence of a proteinaceous substance having a molecular weight of about 180 kDa according to its relative electrophoretic migration rate through detergent-containing polyacrylamide gel is then measured. This substance reversibly binds concanavalin A and is complexed with gamma globulin while in the urine. The gamma globulin complex binds to Staphlococcal protein A. Said proteinaceous substance, when present in detectable amount, is an indicator of bladder cancer.The method of the present invention for diagnosing bladder cancer in a subject may also be described as comprising detection in a urine sample from said subject of a proteinaceous substance having a molecular weight of about 180 kDa and being unreactive with antibodies specifically binding to carcinoembryonic antigen or epidermal growth factor receptor.

Abstract: A dry test strip for the detection of an analyte in a test fluid is disclosed. The test strip comprises a porous detection zone containing a reactant system that can generate a signal in the presence of an analyte. The detection zone further comprises dextran as a barrier to prevent penetration of RBC's into the detection zone.

Abstract: An aqueous wash solution is useful for the detection of herpes simplex virus in a biological specimen. This solution has a pH of from about 9 to about 11.5, and consists essentially of an alcoholamine or a salt thereof and a nonionic surfactant. The solution is used to was uncomplexed materials from a complex of herpes simplex antigen and antibodies thereto. The wash solution can be supplied as part of a diagnostic test kit.

Abstract: A method for separation of erythrocytes from whole blood by diluting the whole blood with saline and introducing low concentrations of soluble salts of trivalent cations into the diluted blood sample.

Abstract: The present invention is directed to a fluorescence polarization assay for benzoyl ecgonine and substituted benzoyl ecgonine compounds in biological fluids, and to a method of making reagents therefor. Specifically, tracers, immunogens and antibodies are disclosed. The tracers and the immunogens are made from substituted benzoyl ecgonine compounds. A fluorescein moiety is included in the tracer, while a poly(amino acid) forms a part of the immunogen. The assay is conducted by measuring the degree of polarization retention of plane polarized light that has been passed through a sample containing antiserum and tracer.

Abstract: An immunoassay procedure for detection of analytes in urine wherein an immunological reaction is conducted in an aqueous phase containing the urine and a filterable immunocomposite containing an inherently colored gold sol particle is formed if the assay is positive. The colored, gold sol particle containing immunocomposite is collected for direct visual observation on a filter element.

Abstract: An enzymic method for assaying protein C by the use of a synthetic peptide substrate, wherein the action of interfering substances against said peptide substrate are specifically inhibited, and a measuring kit for the same.

Abstract: A method for preparing a fecal sample for immunoassay testing comprising the steps of dispersing a sample of from 1 to less than 10 wt. % of a stool sample in an aqueous fecal test solution formulated with one or more preservatives, analyte stabilizing agents and endogenous interference reducing agents. The fecal solids are then permitted to settle to form a liquid phase substantially free from fecal solids, and the clarified liquid phase is removed to provide a test sample free from fecal solids. The fecal test solutions contain suitable stool stabilizers such as buffers and antimicrobial agents, analyte protecting agents such as proteolytic, reductive or oxidative enzyme inhibitors, endogenous assay interfering enzyme inhibitors such as a reducing agent, and non-specific binding inhibitors such as animal proteins. The stool sample should be fresh or be fresh frozen and thawed immediately before dispersion in the buffer solution.

Abstract: A kit and a method for detecting denatured lipoproteins is disclosed. The method comprises: adding to a sample a mixture of (i) an affinity gel with which a monoclonal antibody for apoprotein A-I is coupled and (ii) an affinity gel with which a monoclonal antibody for apoprotein B-100 is coupled; removing apoprotein A-I and apoprotein B-100 in the sample by combining the apoprotein A-I and apoprotein B-100 with the affinity gels; and measuring an amount of lipoproteins in the residue. The kit comprises a mixture of (i) and (ii) as an essential component. The kit and the method is useful for the clinical investigation and diagnosis of abnormal lipid metabolism which can be a cause of heart diseases (e.g. cardiac infarction), diabetes, and various types of atherosclerosis diseases.

Abstract: The present invention is directed to a method for selectively blocking or inactivating glucocorticoid receptors by administering a glucocorticoid receptor blocking effective amount of arsenite or methyl methanethiolsulfonate (MMTS) in a sample. Thus, arsenite and MMTS constitute new, simple, reversible, and inexpensive reagents for assaying glucocorticoid receptors in the presence of other receptors and for eliminating the complications of assaying other receptors in the presence of glucocorticoid receptors.

Abstract: A method for the quantification or qualification of antigens with an immunological solid phase method, for example of the ELISA type, during which the invention seeks to overcome the difficulties with falsely high or falsely positive measurement results.

Abstract: A method is known for the determination of osteocalcin in human serum or plasma, in which a sample of the biological fluid containing the osteocalcin to be determined is incubated, together with a defined amount of an oligopeptide tracer, with a suitable anitbody which binds both the osteocalcin and the tracer. This method has proved to be susceptible to error in that the incubation conditions affected the measured levels, and the osteocalcin levels obtained were often too high. The recognition that the observed errors are attributable to a proteolytic breakdown of the oligopeptide tracer by constituents of the serum or plasma resulted in the technical teaching of the addition of one or more proteolysis inhibitors to the sample of human serum or plasma before or together with the addition of the oligopeptide tracer, specifically either a mixture of an endopeptidase inhibitor with an aminopeptidase inhibitor or, when an N-terminally protected oligopeptide tracer is used, only an endopeptidase inhibitor.

Abstract: A process for the removal of non-specific turbidities in the carrying out of determinations according to the immuno-assay principle, wherein an inorganic boron compound is added to the sample solution in combination with a buffer system.

Abstract: A rapid and specific method and reagent for determining LD-1 isoenzyme in biological fluids by incorporating a chaotropic agent, such as sodium perchlorate, into an LDH assay reagent system. The chaotropic agent produces an immediate inactivation of the LDH isoenzymes containing one or more `M` type subunits while the LTD-1 isoenzyme remains stable and is simultaneously measured.

Abstract: This invention teaches a process for reducing protein matrix effects in assays for serum fructosamine. Blood or blood derived samples are used, and one adds two reagents, one of which reduces interference caused by non-specific reducing substances, the other of which eliminates turbidity. Incubation follows, and then the pH of the sample is adjusted and color forming reagent is added. In one embodiment, the incubation time is only 1-15 minutes. In another embodiment, the first reagent contains peroxidase.

Abstract: Diagnosis of Lyme disease by immunoassay for anti-Borrelia burgdorferi antibodies in the serum of a patient includes adsorption of crossreactive antibodies in the serum with antigen of Acinetobacter calcoaceticus or Treponema phagedensis prior to binding of anti-Borrelia antibodies to Borrelia burgdorferi antigen absorbed onto a solid support. The preferred assay is a flow-through assay using a porous membrane as the support and a dye-loaded liposome conjugated to a goat antihuman antibody for detection. The invention includes a kit of materials for performing the assay.

Abstract: Basement membrane proteins are isolated as functioning proteins in relatively large amounts from human or animal tissues in aqueous solution in the presence of a chelating agent. It is possible to use these proteins to obtain highly specific antibodies which are used for the immunological determination of these proteins.

Abstract: Reagents and an immunoassay for detecting the presence or amount of polychlorinated biphenyls in a test sample. The assay is performed by adding a known concentration of a tracer labeled with a detectable moiety and a known concentration of an analyte-specific antibody to a test sample to form a mixture, incubating the mixture to form labeled tracer-antibody and analyte-antibody complexes, and determining the presence or amount of tracer-antibody complexes formed as a measure of the presence or amount of analyte in the test sample. Reagents provided include tracers, immunogens and an additive compound useful in preventing non-specific binding of the polychlorinated biphenyls to proteins which may be present in the test sample. A kit for performing the assay also is provided.

Abstract: The invention relates to assays utilizing microparticle separation. More particularly, the invention relates to microparticle assays used for confirmation of ligands in a biological sample.

Abstract: A precipitation reagent for use in analytical systems for the determination of hydrophobic analytes in a biological test sample, particularly analytical systems employing specific binding proteins for such analytes. The precipitation reagent precipitates interfering proteins, hemoglobin, and other interfering substances from a biological test sample while, at the same, maintaining hydrophobic analytes in solution and minimizing the denaturation of specific binding proteins, such as, for example, antibodies, which may be present in an immunoassay system. The precipitation reagent comprises a zinc salt, a glycol, and a straight or branced alcohol from about 1 to 4 carbon atoms, and may optionally contain an acid. A preferred precipitation reagent comprises zinc sulfate, methanol and ethylene glycol, and is particularly useful in a fluorescent polarization immunoassay for the determination of hydrophobic analytes, especially cyclosporine.

Abstract: An extraction composition is useful for lysing chlamydial, gonococcal or herpes organisms and extracting detectable antigen from the organisms. In particular, this composition has a high pH (at least about 8) and comprises an alcoholamine which is effective in extracting antigen. It is particularly useful for extracting either or both of the lipopolysaccharide and major outer membrane protein chlamydial antigens.

Abstract: A quantitative immunoassay for a beta-lactam antibiotic in a sample, which may contain open and closed ring forms of the antibiotic, is disclosed. Closed-ring forms of the antibiotic in the sample are converted to open-ring protein conjugates and detected through immunospecific reaction with antibodies raised against the open-ring protein conjugate form of the beta-lactam antibiotic.

Abstract: A flow cytometry method for reproducibly detecting and counting a lymphocyte population of interest in a leukocyte suspension or whole blood sample in which the red cells are subsequently lysed. The suspension (or sample) is combined with a reagent comprising a primary antibody, either native, carrying an attached enzyme or biotin or other label, and a fixative reagent, in either order. Where the enzyme is not attached, an enzyme is coupled specifically to the primary antibody. The fixed suspension is reacted with a color-producing enzyme-cytochemical reagent. The suspension, now including stained and unstained fixed cells, is passed through a flow cytometer and the cells are characterized and counted on the basis of their light-scattering and light-absorbing properties.

Abstract: An immunoassay for trichothecenes that have at least three hydroxyl groups at specified positions is disclosed. It relies on developing antibodies to close trichothecene variants that are missing at least one of the hydroxyl groups, and then using these antibodies to test specimens in which the trichothecene has been converted to the variant (usually to the OAC variant). For example, DON and T-2 tetraol can be assayed for using this invention.

Abstract: A method for the quantitative determination of micro-organisms or pyrogens which comprises introducing a prescribed amount of a test liquor containing micro-organisms or pyrogens into a total carbon analyzer to obtain a total carbon value A, while removing micro-organisms or pyrogens from the prescribed amount of the same test liquor and introducing the resultant into a total carbon analyzer to obtain a total carbon value B, subtracting the total carbon value B from the total carbon value A and determining micro-organisms or pyrogens basing upon the resulting value, and an apparatus suitable for conducting the method.

Abstract: Compounds having detergent properties are disclosed. When a modifying reagent is brought into contact with these compounds, the detergent properties are decreased. These compounds are useful, for example, as solubilizing agents for microbial antigens and/or antibodies and for reversibly wetting hydrophobic surfaces. Accordingly, methods are disclosed for increasing the hydrophilic properties of a material, such as a microbial antigen and/or antibody, the methods generally comprising the steps of contacting the material with the compound having detergent properties and a modifiable group, and modifying the compound with a modifying reagent. Kits are also disclosed for use in accordance with this methodology.

Abstract: The present invention involves a method for producing a substrate useful in a system for the detection of antibodies directed against platelet antigens. This method comprises several steps. A platelet sample of interest is initially treated with an aqueous solution comprising a dialyzable nonionic detergent. This initial treatment is under conditions to solubilize platelet components and produce a platelet lysate. Such conditions may involve treatment of a platelet sample with an aqueous solution comprising nonionic detergent at a concentration between about 0.2% and about 0.5%. Platelet antigens are most preferably solubilized for about 30 min and at about 0.degree. C. in an aqueous solution comprising about 1 mg dialyzable nonionic detergent per mg platelet protein.The partially purified platelet antigens resulting from these manipulations are then preferably affixed to a solid matrix.

Abstract: An assay which is responsive to the levels of anti-p24 antibodies in serum samples is provided which comprises the steps of: (a) forming a test mixture comprising the sample and an antigen solution containing free p24 antigen within a predetermined concentration range: (b) incubating the test mixture under conditions whereby anti-p24 antibodies from the sample, if any, can react with the free p24 antigen to form antibody-antigen complexes; (c) determining the concentration of free p24 antigen remaining in the test mixture after the incubation; (d) determining the concentration of free p24 antigen in the antigen solution; and (e) calculating the difference between the concentration of free p24 antigen in the antigen solution and the concentration of freee p24 antigen in the test mixture after the incubation.The difference calculated in step (e) represents the "binding capacity" of the serum sample for p24 antigen.

Abstract: There is provided a biological diagnostic assay system wherein a phenoxy-substituted naphthalene compound, such as phenoxynaphthalene sulfonate, or a salt thereof, is utilized to prevent plasma proteins such as serum albumin from binding to other components of the assay and/or to displace plasma proteins which have become bound to other components of the assay.

Abstract: A material and method are described to reduce the non-specific binding of a labelled material, wherein the labelled material comprises a fluorochrome and a specific binding pair member. The invention is particularlly suited to the lysis of peripheral blood wherein a labelled material is used to identify and detect cell populations within a sample of blood.

Abstract: A method for preparing a fecal sample of immunoassay testing comprising the steps of dispersing a sample of from 1 to less than 10 wt. % of a stool sample in an aqueous fecal test solution formulated with one or more preservatives, analyte stabilizing agents and endogenous interference reducing agents. The fecal solids are then permitted to settle to form a liquid phase substantially free from fecal solids, and the clarified liquid phase is removed to provide a test sample free from fecal solids. The fecal test solutions contain suitable stool stabilizers such as buffers and antimicrobial agents, analyte protecting agents such as proteolytic, reductive or oxidative enzyme inhibitors, endogenous assay interfering enzyme inhibitors such as a reducing agent, and non-specific binding inhibitors such as animal proteins. The stool sample should be fresh or be fresh frozen and thawed immediately before dispersion in the buffer solution.

Abstract: A method for producing a binding assay device composed of antigens on a cellulose nitrate, cellulose nitrate/acetate or similar solid phase is described. The method involves applying to a solid phase a small amount of an allergen composition, or a pretreated allergen composition, containing a certain concentration of allergen and drying the solution. The device is used by contacting a patient test sample to the immobilized allergen and determining whether or not the test sample contains IgE antibodies for the allergen.

Abstract: This invention is directed to a ligand-receptor assay for determining the presence or amount of at least one target ligand, capable of competing with a ligand analogue conjugate for binding sites available on a ligand receptor, said ligand analogue conjugate comprising at least one ligand analogue coupled to a signal development element capable of emitting a detectable signal, in a fluid sample suspected of containing said target ligand, comprising the steps of:a. contacting said fluid sample with ligand analogue conjugate and ligand receptor to form a reaction mixture, the relative amounts of ligand analogue conjugate and ligand receptor being such that in the absence of target ligand, and subsequent to substantially equilibrium binding, substantially all of the ligand analogue conjugate is bound to ligand receptor;b. detecting the unbound ligand analogue conjugate;c. relating the detectable signal to the presence or amount of target ligand in the fluid sample.

Abstract: An attenuator is included in a reagent medium of a luminescent specific-binding assay to suppress undesirable extraneous light. In one such assay, an analyte in a sample is reacted with a specific binding partner attached to a solid surface, forming an immobilized pair at the surface. One member of the immobilized pair is then allowed to react with a specific binding partner previously conjugated with one component of a luminescent reaction system, and the remaining components are provided in the reagent medium. The resulting light emitted in the luminescent reaction is recorded on photographic film or other photodetector as a measure of the presence and quantity of the analyte in the sample.