Abstract: This invention provides methods for the detection of a target genetic material having a desired base sequence or gene. Also disclosed are methods for the detection of mutations. Also provided are components for use in such methods.The methods are based upon techniques which utilize two labeled single stranded polynucleotide segments which are complementary to the same or the opposite strands of the target genetic material. The methods of the invention result in the formation of a double hybrid and/or multihybrid.

Abstract: Assay methods are provided for determining an analyte in a sample suspected of containing the analyte. The method is carried out using a composition that includes a conjugate of a first sbp member with a particle. A luminescer is reversibly associated with a nonaqueous phase of the particle. Where the first sbp member is not complementary to the analyte, a second sbp member that is capable of binding to the first sbp member is employed. Unbound conjugate is separated from conjugate that is bound to the analyte or to the second sbp member. A reagent for enhancing the detectability of the luminescer is added and the light emission of the luminescer acted on by the reagent is measured.

Abstract: The invention relates to an immunological complex of two antibodies of a first animal species, e.g. mouse antibodies, which have been conjugated to form a cyclic tetramer with two monoclonal antibodies of a second animal species, e.g. rat monoclonal antibodies, directed against the Fc-fragment of the antibodies of the first animal species.Preferably, the complex is bifunctional, that is to say, that it contains two different antibodies of the first animal species, one of which is preferably directed to a detectable substance, such as an enzyme, e.g. peroxidase, and the other is directed to any desired antigen. A tetrameric complex of this kind may be used as a labelled antibody in immunoassays.Further, the invention relates to processes for preparing the complexes and for using the complexes in immunological reactions. Finally, the invention relates to drugs containing a complex according to the invention, as the complexes may be used to direct certain active principles to target cells.

Abstract: A method is provided for detecting the presence of C. difficile toxin A. Stool or other appropriate specimen is contacted with a reagent containing an available non-reducing galactose-alpha-1-3-galactosyl structure, which is a specific receptor for toxin A. The reagent may be intact cells, cell membranes, membrane fractions containing the toxin A receptor structure, glycoconjugates, as well as the purified toxin A receptor per se. Binding of toxin A is determined by conventional assay techniques. The method may also be used to isolate and purify toxin A. Conversely, immobilized toxin A is used to detect, isolate, or purify biological materials of interest containing the receptor structure.

Abstract: This invention provides a means for determining the concentration of any of a wide range of antibody or antigen molecules with a high degree of specificity, accuracy and sensitivity. Antigen or antibody concentration is determined by effecting an agglutination reaction in a liquid medium and determining the cluster size distribution of agglutinated particles by optical pulse particle size analysis. The measured cluster size distribution then is compared with a standard quantitative relationship between the cluster size distribution and concentration of the antigen or antibody being tested. By this means one may specifically ascertain the absolute concentration of the antigen or antibody in question in the sample being analyzed. In addition to detecting antigen or antibody molecules, the process of this invention can be used to determine the concentration of any substance capable of specifically promoting or inhibiting an agglutination reaction such as viruses, white blood cells or the like.

Abstract: A method for detecting the presence in a human of an integral membrane calcium-binding protein associated with essential hypertension which comprises isolating tissue from a human, treating the tissue to obtain integral membrane proteins, contacting the proteins thus obtained with a first antibody molecule which binds to the integral membrane calcium binding protein to form an detectable protein-antibody complex, and detecting the complex so formed.Further, methods for quantitatively determining the amount of an integral membrane calcium-binding protein and the messenger RNA encoding said protein, diagnostic methods for identifying individuals predisposed to essential hypertension, and protein and messenger RNA associated with said hypertension.

Abstract: A historical time series analysis is obtained from a single sample of blood by measurement of hemoglobin and an altered hemoglobin, for example, glycohemoglobin, on a cell-by-cell basis. The results are compared by ratio of glycohemoglobin to hemoglobin on a cell-by-cell basis. Since the alteration of hemoglobin is continuous and irreversible, the ordered set of ratios represents a time-series of the cells, and any deviation from normal will be readily apparent A time-series can also be derived for other constituents, the time-series representing historical data of the other constituents in the person from whom the blood sample was taken.

Abstract: A method of detecting the presence and/or amount (concentration) of an analyte in a sample is provided by forming a reaction medium containing (1) a sample; (2) a plurality of particles having a binding pair member bound to their surfaces; and (3) a monovalent complementary partner to the binding pair member to which is bound an analyte mimic or analyte binding partner; and detecting the presence or amount of agglutination of the particles in the reaction medium. In some cases a polyvalent receptor capable of binding to the analyte (and to the analyte mimic, if present) is also present in the reaction medium.

Abstract: A method for immunological assay of IgM antibodies against at least one said antigen in a liquid sample containing at least IgM and IgG antibodies, such as a biological fluid, comprising reaction of said liquid sample and of antibodies against IgG, and addition to the so obtained reaction product of the said antigen bound to finely divided particles and of a free said antigen, the amount of IgM being inversely proportional to the amount of unagglutinated finely divided particles.

Abstract: Immunoassay which can be performed within a short period of time and a reagent composition for such immunoassay. The reagent composition contains complement and cells which contain quantitatively determinable substance therein. Antibody or antigen corresponding to antigen or antibody to be determined in a sample is bound on the outer surfaces of the cell membranes, which can be lysed by the complement upon activation thereof. In the immunoassay, a sample containing antigen or antibody to be determined is mixed with the reagent composition. The complement is activated by the resultant antigen-antibody complex. The cell membranes are lysed by the complex and the determinable substance in the cells is liberated from the cells. The amount of the liberated substance is determined.

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: Compounds and methods are disclosed for reversibly aggregating particles suspended in a liquid medium. The method comprises combining the liquid medium containing the particles with a polyionic polymer capable of aggregating the particles under conditions suitable for such aggregation. Thereafter, the particles are contacted with a chemical reagent capable of cleaving the polyionic polymer under conditions sufficient to reverse the aggregation. Optionally, magnetic particles are added to the liquid medium in the present method under conditions for non-specific binding and the medium including the aggregates is subjected to a magnetic field gradient to separate the aggregates from the medium. The compounds of the present invention are polyions. The aggregation of the particles is reversible upon contact with chemical agents which cleave at least some of the bonds within the polyionic polymer.

Abstract: A number of naturally occurring antibodies to human erythrocyte surface antigens are capable of combining with their specific antigens (for example, Rhesus factor), but are not capable of producing visible hemagglutination. Also, the sensitivity of many diagnostic methods, such as in human blood typing, depends upon cell agglutination.The present invention provides liposome-protein conjugates, especially useful for hemagglutination assays, having an enhanced agglutination capacity with respect to antibody from which the conjugates are derived.

Abstract: The invention is directed to a method for the diagnosis of a malady in a subject by observing a degree of reaction between all blood cells: red blood cells, leukocytes and platelets; in the subject's blood with a foreign entity having a predetermined relationship with the malady being diagnosed. The test includes comparing amounts and sizes of red blood cells, leukocytes and/or platelets in a control sample and at least one test sample. The test sample includes a portion of the subject's blood and the foreign entity being tested.

Abstract: Disclosed herein is a method for examining cells, comprising subjecting the cells to an antigen-antibody reaction treatment, then measuring a pattern of the electrophoretic mobility of the cells and comparing the electrophoretic property of the cells under examination with the electrophoretic property of standard cells.

Abstract: An improved assay for an ATL virus antibody in a specimen, in which an ATL virus antigen is added to the test specimen and used as a comparative specimen. This improved method selectively assays ATL virus antibodies, and, thus, is useful in preventing and treating adult T cell leukemia.

Abstract: A method is provided for determining the presence in a sample of a member of a specific binding pair ("sbp member") consisting of ligand and its homologous receptor. The sample is combined in an aqueous medium with (1) a complementary sbp member wherein at least the sbp member or the complementary sbp member is bound to the surface of a cell and (2) a fuorescent agent capable of being incorporated into the cell. The presence of the sbp member is indicated by a change in fluorescence of the unseparated cell suspension as a result of agglutination of the cells.The present invention has particular application to blood typing, for example, for the determination of the presence of blood group antigens A, B, AB, O, and D (Rh.sub.o) and antibodies to such antigens.

Abstract: A method and composition for detecting analyte moieties by means of a signalling moiety capable of aggrandizement are disclosed. The signalling moiety can be attached to or not attached to the analyte moiety/analyte-specific moiety complex. The signalling moiety can be viable or non-viable. The methods disclosed herein provide a sensitive assay for the detection of a wide range of different analyte moieties.

Abstract: A blood fluid composition for use in a complement-mediated cell lysis system. The composition includes a blood fluid, which may be either a serum source of complement, analyte-containing serum, or both, and lipid vesicles capable of reducing the extent of non-specific cell lysis produced when the blood fluid is added to lysable target cells in the system. The vesicles are present in an amount which increases the ratio of ligand-specific to non-specific cell lysis in the system at least 2-fold and preferably 4-fold or more over that achievable in the system in the absence of the vesicles.

Abstract: A viral lysis immunoassay system and method. The system includes hemolytic particles carrying non-viral, anti-analyte molecules, lysable target cells which are devoid of surface molecules capable of binding to endogenous viral surface molecules, and foreign binding molecules added to the target cells. The binding molecules, which may be either analyte molecules or analyte-related molecules attached to the target cell surfaces, function to bind the particles to the cells, to initiate cell lysis and the release of encapsulated reporter molecules from the cells. The analyte to be assayed may be one adapted to bridge the virus particles to analyte-related molecules carried on the cell surfaces, or one which competes with target-cell molecules for binding to the particle anti-analyte molecules.

Abstract: Methods are provided for the detection of proximal convoluted tubule causing diseases or kidney harmful drug monitoring by detecting the presence of shed normal proximal tubule associated antigens in a body fluid such as urine. The preferred embodiment employs an ELISA sandwich format wherein one monoclonal antibody specific for a first epitopic site on said antigen is immobilized on a solid phase and a second antibody, specific for a second epitopic site on said antigen is directly or indirectly labeled.

Abstract: A process for the detection and assay of a biological substance by erythroadsorption by immobilizing a substance having a binding affinity for the biological substance to be assayed; incubating the immobilized substance with a liquid medium containing the biological substance to be assayed, forming a fixed substance; incubating the fixed substance with a coupling product comprising a specific ligand and a second ligand which binds with erythrocytes; adding erythrocytes; and determining the amount of erythrocytes bound to the coupling product. The determination of the amount of bound erythrocytes can be made visually with the naked eye or determined by lysis of the erythrocytes bound to the coupling product and quantitatively measuring the released hemoglobin. The specific ligand can be an antibody or an antigen and the second ligand capable of coupling erythrocytes can, for example, be red blood cell antibodies.

Abstract: A process of immunoassay of a selected substance in a liquid sample containing one or more other non-selected substances, each of these substances being comprised of at least two structurally different chains, the process comprising among others addition of a dissociating agent, such as for example a proteolytic enzyme, to the sample, mixing of the so-obtained liquid, inactivation of the dissociating agent, addition of antiserums, and determination of the amount of this selected substance in the liquid sample.

Abstract: A method of measuring an infectious disease antibody such as syphilis IgM, which comprises treating immunoglobulins of a sample with an anti immunoglobulin antibody sensitized on carrier particles and an antigen of the infectious disease sensitized on carrier particles. According to the method of the invention, the specific antibody of the specific infectious disease such as syphilis IgM can easily and exactly be measured. This method is useful for the judging the stage of infectious disease and watching the results of treatment.

Abstract: A novel method for the diagnosis of Acquired Immune Deficiency Syndrome (A.I.D.S.) is disclosed, involving the use of immunocytoadherence (rosette inhibition) techniques. The method is based on the discovery that the lymphocytes of A.I.D.S. patients are unusually resistant to antithymocyte serum, and that the plasma of A.I.D.S. patients is capable of conferring this resistance to normal lymphocytes. Accordingly, the diagnostic method involves performing rosette inhibition tests on the patient's lymphocytes or on lymphocytes from a healthy donor after treatment with the patient's plasma. Any observable lessening of inhibition in comparison with a control is related to the presence of A.I.D.S.

Abstract: An immunoassay of the type which utilizes the hemolysis of microcapsules such as red blood cells. Microcapsules which can be lysed by complement activity, which contain an optically determinable substance, on whose surfaces an antibody to be quantified is bound are prepared. The microcapsules, a test sample containing the antigen or the antibody to be quantified, and complement are mixed to react each other. Thereafter, an optical measurement is conducted at different wavelengths for the reaction mixture which is still suspending the intact microcapsules.

Abstract: Methods are provided for rapidly determining a number of parameters in a few determinations. Particularly, the method is applicable to blood typing, determining the blood type as to the ABO and Rh type, as well as the determination of isoantibodies to the antigens. The method employs fluorescent particles having a plurality of fluorescers, where the presence or absence of light emission of a particular wavelength can be determined.

Abstract: A specific binding assay composition and method for determining a ligand in a sample are disclosed. The composition comprises (a) a binding partner for the ligand; (b) a detection system which has at least two components; (c) a selectively accessible vesicle having a surface-incorporated ligand or ligand analog and a first component of the detection system therein; (d) a substance which modifies vesicle accessibility in response to binding of surface-associated ligand or ligand analog and binding partner; and (e) at least one additional component of the detection system which is reactive with the first component to produce a detectable response which is reduced by association of the binding partner and vesicle modifying substance with the variabily accessible vesicle. A decrease in signal is measured upon reaction as compared to the signal of unreacted vesicles.

Abstract: A plurality of ampules, individually formed, filled with reagents, sealed with a pane and marked with indicia for reagents to be mixed with blood in performing tests to determine blood type groups is disclosed. A receptacle for a blood sample which conveys the blood sample to the ampules and which accomodates a plurality of the ampules is also disclosed along with a means for rupturing each pane.

Abstract: Early pregnancy can be determined by detection of a protein in milk, which protein is associated with the placental membrane. The protein is characterized by having an approximate molecular weight of 47,000 to 53,000 and an isoelectric point of from about 4.0-4.4.

Abstract: An assay utilizing receptor or antibody sensitized liposome particles which have ATP encapsulated therein. The ATP is released by lysing the liposomes, and detected by means of luciferin-luciferase reagent and a luminometer. The assay provides a very sensitive process for detecting the presence of analytes such as antigens and DNA probes.

Abstract: In order to produce a more well defined interface between adjacent cell layers in a centrifuged sample of anticoagulated whole blood, a material which will bond one group of cells together is added to the blood sample prior to centrifugation. The bonding material must produce a high strength bond between one group of cells, but not effect the other cell types. The material is added prior to centrifugation of the blood sample. An example of a bonding agent is a monoclonal antibody.

Abstract: A method for the quantification of an antigen, antibody, or antigen-antibody complex in a sample solution involving measuring the results of an immunochemical agglutination reaction or an agglutination inhibition reaction by spectrophotometry, wherein after the initiation of the agglutination between the antigen, antibody, or antigen-antibody complex to be determined and sensitized carrier particles to which a substance specifically bindable to the said antigen, antibody, or antigen-antibody complex is bound, a fixing compound is added to fix aggregates formed by the agglutination. A measuring reagent kit or pack includes all the components for use in carrying out the above method.

Abstract: A process for the detection of a biological substance immobilized on a support including(1) incubating, after washing, the substance immobilized on a support with the product of coupling of a specific ligand with a ligand capable of reacting with erythrocytes;(2) adding erythrocytes;(3) immersing the whole in a solution of a fixing agent and(4) measuring the erythroadsorption, after having turned the support over in order to permit the removal of the red cells which have not reacted.

Abstract: The invention relates to techniques for isolating from a mixed population of cells disired living cells either producing and releasing a particular product or having a characteristic molecule on their surface. The isolation techniques depend upon the localized interaction between the product (or molecule) and other agents added to the system such that distinguishable conditions can be caused to occur (or not occur) only in the immediate vicinity of desired cells which produced and released the product or which contain the molecule on their surface.

Abstract: A preparation for the detection of chlamydial infections using lipopolysaccharide of Re-lipopolysaccharide mutants of gram-negative bacteria. The lipopolysaccharide preparation is used in the production of group-specific antibodies to chlamydiae for diagnostic purposes or for the demonstration of antibodies to chlamydial group antigen in specimens.

Abstract: Two monoclonal antibodies (3-3 and 3-40) were produced which identify two new leukemia associated antigens. Both antibodies reacted with most cell lines derived from patients with T lymphoblastic leukemia (T-ALL), but were not detected on suspensions of normal hematopoietic cells, including thymocytes, by cytotoxicity, absorption of indirect immunofluorescence assays. Analysis of fresh leukemic cells indicated that mAb 3-3 only reacted with T-ALL cells, while mAb 3-40 reacted with some non-T non-B ALL cells and a few acute myelocytic leukemia (AML) cells, as well as T-ALL cells. The 3-40 antigen was also found histopathologically in frozen sections of several normal tissues, including the epithelial cells and a few lymphoid cells of the thymus, and some malignant tissues, while the 3-3 antigen was not found in any tissue studied. A "double absorption" assay provided additional serological evidence the the two antibodies identify different antigenic determinants.

Abstract: A method and system for detecting and preferably measuring the presence of an activated complement complex in a sample is discussed. The presence of such an activated complex is indicative of complement pathway activation and includes a first complement component and a second complement component. The method uses a first binding agent specific to the first complement component and a second binding agent specific to the second complement component which when bound with the complex forms an aggregate. The second specific binding agent includes a label whose presence is used to detect and measure the amount of aggregate and therefore activated complex in a sample. An assay system and aggregate for use in an assay system are also discussed.

Abstract: An enhanced agglutination assay method for determination of a multivalent analyte is disclosed. Analyte is added to agglutinatable particles coated with anti-analyte molecules to produce particle agglutination. The extent of agglutination is enhanced by mixing the particles and analyte with an analyte-binding reagent composed of lipid bodies. The reagent bodies act by promoting multiple analyte bridge connections between individual bridged particles and a reagent body. Also disclosed is a kit containing such particles and reagent.

Abstract: The present invention relates to the carrying out of immunochemical reactions in a test tube having a bottom in the shape of an inverted prism or truncated prism, in such a way that the lower edge of the tube forms a strip having a length which is at least equal to one-quarter of the distance between two opposite walls, or where appropriate to a quarter of the diameter of the test tube, as a result of which both the sensitivity and the readability are improved.

Abstract: A reagent for determination of human blood coagulation factor XIII for reversed passive hemagglutination reaction which comprises sensitized erythrocytes prepared by sensitizing animal erythrocytes with specific anti-human factor XIII antibody, and a kit using the reagent are disclosed.

Abstract: An immunoassay by which a plurality of kinds of antigens or antibodies in one test sample may be simultaneously quantified. Each of a plurality of kinds of microcapsules is first provided for each kind of a plurality of antigens or antibodies to be quantified such that the microcapsules bind an antibody or antigen specific to one kind of the plurality of antigens or antibodies to be quantified. The microcapsules are formed of a membrane capable of being lysed by the complement activity, and each kind of microcapsules contains therein a substance that is quantifiable and does not interact with another quantifiable substance contained in other kinds of microcapsules. The plurality of kinds of microcapsules are mixed with a test sample and complement, and the quantifiable substances are released from the microcapsules upon lysis of the microcapsules by the complement activity. The quantifiable substances are then quantified.

Abstract: A reagent and merchandizing kit for use in the diagnosis of syphilis by hemagglutination of an anitgen obtained from a culture of pathogenic Treponema pallidum Nichols and which is sensitized on carrier particles in the presence of antibody wherein said antigen is substantially devoid of proteinic fractions of said culture having a specific gravity of less than 1.01, and methods of preparing the same.

Abstract: Aqueous formulation containing monoclonal antibodies active against cell-bound antigens, particularly human red cell antigens, the said formulation containing a soluble salt in a concentration of not less than about 200 mmoles/1.

Abstract: A method and serological reagent for determining antigens or antibodies with enhanced sensitivity and specificity techniques acts in an interdependent manner in one diagnostic unit of two separately acting components. This action can be simultaneous or consecutive and the combined effect inhibits "lectin-like" factors or "unspecific antibodies" and detects specific antibodies or antigens.

Abstract: A method of forward blood grouping is presented wherein known antibodies are attached to a solid surface and the red blood cells presented for immunological reaction are activated with a proteolytic enzyme. A reverse blood grouping procedure utilizes synthetic or purified antigens which are attached directly to a solid surface. The surface is then contacted with an unknown blood component to permit antibodies to undergo immunological reaction with the previously attached antigens. A solution of red blood cells is used as the indicator. A method of performing a major crossmatch utilizes anti-human immunoglobulin that is attached to a solid surface which is then contacted by the two blood components to permit an antigen-antibody interaction of red blood cells and antibodies. The antibody sensitized cells will immunologically adhere to the solid phase.

Abstract: A number of naturally occurring antibodies to human erythrocyte surface antigens are capable of combining with their specific antigens (for example, Rhesus factor), but are not capable of producing visible hemagglutination. Also, the sensitivity of many diagnostic methods, such as in human blood typing, depends upon cell agglutination.The present invention provides liposome-protein conjugates, especially useful for hemagglutination assays, having an enhanced agglutination capacity with respect to antibody fromThe invention described herein was made in the course of work under a grant or award from the Department of Health and Human Services.

Abstract: Methods and compositions are provided for performing an assay on whole blood samples. The method is for a determination of an analyte which is a member of a specific binding pair (sbp) consisting of ligand and homologous receptor. The method involves a binding agent for the red blood cells in such sample, a solid bibulous element to which is bound at least one sbp member, and a signal-producing system. The method comprises combining the whole blood sample, the binding agent, and none or, where appropriate, one or more members of the signal producing system. The medium is next contacted with a portion of a solid bibulous element to which is bound one of the members of the specific binding pair to allow the medium to traverse such element (immunochromatography). The solid bibulous element is contacted with any remaining members of the signal producing system. Signal resulting from the signal producing system is detected and is related to the amount of the analyte in the sample.

Abstract: This process involves the use of benzoquinone as cross-linking agent in large excess compared with the substance to be activated. The activation reaction is realized in a homogeneous liquid medium. Said process making it possible for example to couple antibodies to enzymes for determining or detecting said antibodies. The process can be used for the determination of antitetanic antibodies in human serum.

Abstract: A direct particle agglutination immunoassay for assaying an antigenic substance (Ag) in a fluid. The immunoassay is the type which comprises:(a) contacting the fluid with an antibody (Ab) coated particle (P) to agglutinate the Ab coated particles; and(b) detecting the presence of agglutination.The immunoassay is characterized in that the fluid is contacted with at least one additional entity selected from a group consisting of at least one different type of antibody (Ab.sub.a) coated particle (P.sub.1) and at least one different type of antibody (Ab.sub.b). The P.sub.1 is selected from a group consisting of P, at least one different particle (P.sub.2), and mixtures thereof (P and P.sub.2). Each type of Ab.sub.a -P.sub.1 and Ab.sub.b has a lower average affinity constant (K) for Ag than the K of Ab-P and the additional entity is present in an amount sufficient to avoid a high antigen false negative effect.Also, a reagent of the type comprising Ab-P.