Abstract: Immunoassays of psychoactive drugs including psychotomimetic drugs, narcotic drugs, and tetrahydrocannabinols and treatment methods based on the antigenic properties of protein conjugates of these drugs. These methods are based upon treating the psychoactive substances as haptens and utilizing their protein conjugates to produce antibodies to the psychoactive materials themselves. The immunoassay methods include both agglutination and agglutination-inhibition reactions. The treatment methods include treatment or both exogenous, administered drugs (such as cannabinols, LSD, heroin and morphine) and endogenous substances (such as N,N-Dimethyltryptamine and 5-Methoxy-N,N-Dimethyltryptamine by active immunization and also passive immunization.

Abstract: Disclosed are materials and methods for detecting biomolecules in samples employing transponders having memory elements associated with particle(s) used as a solid phase in art assay, and information pertinent to the assay is encoded on the transponder memory elements. A dedicated read/write device is used remotely to encode or remotely to read the information encoded on the transponder memory elements. The invention can be used in direct or competitive ELISA-type assays, or in multiplex assays for the simultaneous assay of several analytes.

Abstract: A method is provided for the preparation of semisolid particles or cells that have been chemically derivatized resulting in hydrazide functionalities covalently incorporated onto cell surface membrane molecules of the semisolid particles or cells, as well as a method for determining the levels of hydrazide groups on the particles or cells. The method involves the preparation and analysis of red blood cells chemically derivatized in such a manner that hydrazide functionalities have been covalently incorporated onto the cell membrane molecules, the preparation of hydrazine derivatized cells and the use thereof of the novel cells in agglutination reactions to promote the attachment of antigens and antibodies to the hydrazine derivatized cells without loss of antibody activity or the blocking of the etitope of interest.

Abstract: HIV-1 peptides having at least one point mutation between position 593 and 611 of the HIV-1 gp160 amino acid sequence. The point mutation either is at position 604 or 610, or both positions. Immunoassays which utilize these peptides are provided, as well as, diagnostic test kits which contain these peptides.

Abstract: A process for preparing a purified syphilis antigen which comprises adsorbing an extract originated from Treponemda pallidum on a hydroxyapatite gel, followed by elution, while an aqueous medium is used.

Abstract: A composition is described which is useful in the assay of folate from a hemolysate prepared from red blood cells using non-radiolabeled assay techniques such as fluorometric, colorimetric, enzymatic or chemiluminescent assay methods. Methods to ready hemolysates for the assay of folate are also described.

Abstract: Drug-dependent antibodies that bind to granulocytes, erythrocytes, platelets or membrane proteins derived from these cells, in the presence of a drug, but not in its absence, can be detected using a sensitive assay. Detection of the drug-dependent antibodies permits diagnosis of cytopenia mediated by the drug.

Abstract: Human red blood cells with antigenic determinants recognized by unexpected alloantibodies are mixed with serum from the potential recipient. Following incubation, multivalent monoclonal IgM antibodies specific to the antigenic determinant present on the test red blood cells are added. If the recipient's serum is negative for an unexpected antibody which would bind to the antigens present on the test red blood cells, then the multivalent antibody will agglutinate the red cells. Conversely, if an unexpected antibody specific to those determinants is present in the recipient's serum, hemagglutination will be inhibited.

Abstract: For the detection of proteins containing phosphorylated tyrosine the sample of a body fluid is incubated with at least two receptors R.sub.1 and R.sub.2 whereby a change in signal is produced by binding of at least the receptors R.sub.1 and R.sub.2 with the substance to be detected in the sample solution and in which in each case one of the two receptors contains an antibody or its derivative capable of specifically binding to the protein to be detected and the other receptor contains an antibody or its derivative capable of specifically binding to phosphorylated tyrosine and the signal change caused by the binding is determined in the sample.

Abstract: A method is disclosed for separating a substance from a liquid medium. The method comprises combining the liquid medium containing the substance with magnetic particles under conditions for non-specific chemical binding of the magnetic particles. Thereafter, the medium is subjected to a magnetic field gradient to separate the particles from the medium. The preferred non-specific binding is achieved as the result of charge interactions between the particles usually by means of a polyionic reagent. The method of the invention has particular application to the separation of cells and microorganisms from aqueous suspensions and also to the determination of an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp). The sample is combined in an assay medium with magnetic particles and a sbp member complementary to the analyte.

Abstract: A method for the non-radioactive determination of circulating red cell volume, total blood volume and red cell survival. Red blood cells are biotinylated and injected into the subject for dilution in the subjects total blood volume. A diluted sample is extracted and incubated with a label, either a radionuclide or a fluorescent moiety complexed with avidin or streptavidin. Detection of the label, as for example by gamma counting in the case of a radionuclide or fluorescence activated cell sorting in the case of a fluorescent moiety, allows calculation of the red cell volume and therefrom, total blood volume. Sequential measurements are possible with this method. Aspects of the method include enhanced biotin labeling and separation of cells by density gradient means.

Abstract: The present invention relates to immunoassay methods for detecting and measuring the amount of an analyte in a sample by means of generic anti-hapten antibodies. Also disclosed are multi-analyte immunoassay methods. Reagents, devices, and kits using the anti-hapten antibodies are also disclosed. The present invention also relates to immunoassay methods for detecting and measuring the amount of an analyte in a sample by means of a dual antibody format.The present invention also relates to dyed erythrocytes, preferably fixed, which are coated with antibodies. Also disclosed is the use of these dyed erythrocytes in agglutination assays to detect and measure the presence of an analyte in a sample. The analyte can be a hapten, an antigen, or an antibody. Also included are agglutination assays, compositions and kits using these dyed and coated erythrocytes.

Abstract: A suspension of inert particles is prepared in an aqueous solution, to which an antibody or an antigen and a carrier-bound antigen or antibody, respectively, are added in any desired order. After centrifuging, the positive, weakly positive, or negative reaction can easily be recognized on the basis of a simple pattern.

Abstract: A method of determining the presence and quantity of an analyte of interest in a particulate-containing sample is disclosed, as is a construct for use in the method. The method is particularly useful for determining an analyte in whole blood and in fermentation suspensions. The construct is comprised of a first moiety, which is a particulate-binding moiety and a second moiety, which binds the analyte of interest.

Abstract: A vessel for conducting blood cell agglutination assays is disclosed. A barrier retains reactants in an upper chamber during incubation, then, in response to a force, permits reagents to enter a lower chamber containing a matrix for separating agglutination.

Abstract: An article for performing solid-phase immunological assays and methods for performing assays utilizing such article. A solid-phase support suitable for immunological assays is stained with an organic dye that has a net-positive charge and a hydrophobic aromatic ring structure. An immunologically reactive component having a net-negative charge, such as a whole cell, is securely immobolized by the dye to the support through noncovalent interactions. A sample of biological fluid may be added to the article to which the immunologically reactive component has been immobilized to effect binding of antigens or antibodies in the sample which are specific to the immobilized component. The presence of the antigen-antibody complex may then be determined by any known means.

Abstract: Methods and cards for detecting an antigen or antibody in a fluid sample are disclosed. The fluid sample is added to a microreaction vessel containing a slurry suspension of inert particles and a binding partner to the antigen or antibody to be determined. The fluid sample is added to the micro reaction vessel, with one of the antigen and antibody binding partners being carrier bound. The vessel contents are centrifuged, to cause the binding partners to contact each other to form an optically detectable binding complex. The location of the optically detectable complex is observed to determine the presence of the antibody.

Abstract: In an erythrocyte agglutination assay, the agglutination reagent comprises at least one erythrocyte binding molecule coupled to at least one specific analyte binding molecule wherein the erythrocyte binding molecule does not cause agglutination when incubated with erythrocytes in the absence of analyte (in the case of a direct assay) or analyte binding reagent (in the case of an indirect assay). Preferably, the erythrocytes are endogenous to the blood sample to be tested, that is, a whole blood sample is assayed. Mixtures of conjugates and conjugates of analyte analogues with erythrocyte binding molecules may also be used as agglutination reagents. The reagents and their use in direct or indirect assays is disclosed.

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: This invention relates to a method for the immunoassay of AZT (3'-azido-3'-deoxythymidine), also known as zidovudine, in biological fluids such as serum, semen, plasma and urine, as well as other body fluids. The invention also includes (1) various novel analogs of AZT useful in preparing immunogens for antibodies to AZT and in preparing labeled AZT, (2) immunogens for antibodies to AZT, (3) monoclonal and polyclonal antibodies to AZT, (4) labeled AZT analogs and (5) diagnostic test kits for the immunoassay.

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: For the determination of a specifically bindable substance by incubation of the sample solution with at least two receptors R.sub.1 and R.sub.2, whereby R.sub.1 and R.sub.2 are capable of binding to each other and R.sub.1 is capable of specific binding to the substance to be determined and measurement of the agglutination which occurs in the reaction, a conjugate of one partner of a specific binding pair P and a component K capable of specific binding to the substance to be determined is used as the receptor R.sub.1 and a receptor which has at least two binding sites for P is used as R.sub.2.

Abstract: A method of detecting target antibodies or antigens by reaction with specific binding partners thereto is disclosed. One of the target or the binding partner is bound to a carrier, and the other is unbound. The complex between the target and the binding partner, with one being carrier-bound, forms an optically detectable binding complex. A microreaction vessel having an upper portion, a transition portion and a lower portion is utilized, wherein the upper portion has a greater diameter or width than the lower portion, and the transition portion is situated between the upper portion and the lower portion, and is funnel shaped. The microreaction vessel contains a slurry or suspension of inert particles, and unbound target or binding partner thereto. A solution of the carrier bound target or binding partner thereto is added to the vessel, which is then centrifuged to produce an optical determination of the target.

Abstract: The invention relates to a method of preparing a reagent for the determination by hemagglutination of antibodies to bacterial toxins. According to this method erythrocytes are treated with glutaraldehyde and then with the bacterial toxins in the presence of glutaraldehyde without a wash step. The reagent thus obtained is further treated with a reagent for blocking aldehyde groups.

Abstract: The invention includes an agglutinant complex which is useful for the investigation of the antigens present on erythrocytes, and which results from affinity couplings between nonagglutinant IgG type antibodies specific for the antigen to be identified, a protein capable of binding to at least two sites on the Fc part of antibodies and an anti-immunoglobulin antibody or its fragments.

Abstract: A detecting reagent for an antiplatelet antibody, comprises a carrier particle, and a platelet antigen component immobilized on a surface of the carrier particle. The reagent particles do not agglutinate with each other, contrary to the platelets subjected to natural agglutination, probably because the platelet membrane antigen component immobilized on the carrier particle is solubilized in advance.

Abstract: This invention provides methods and compositions for the detection of a target genetic material having a desired base sequence or gene. Also disclosed are methods and compositions for the detection of mutants. The methods and compositions are based in part upon techniques which utilize two labeled single-stranded polynucleotide segments which are complementary to different portions of the genetic material. The methods and compositions of this invention result in the formation of a double hybrid and/or multihybrid, and can furthermore be employed in an attached system, i.e., a matrix-bound entity capable of binding to a polynucleotide probe entity.

Abstract: The type of a blood specimen is determined by adding a portion thereof, either a diluted solution of its red cells or a portion of its plasma, to each of a multiplicity of wells in a transparent tray, by adding a blood type specific reagent to each of the wells in the tray, different reagents being added to different ones of the wells of the tray, periodically tilting the tray at a substantial angle of at least approximately 50.degree.

Abstract: The present invention relates to immunoassay methods for detecting and measuring the amount of an analyte in a sample by means of generic anti-hapten antibodies. Also disclosed are multi-analyte immunoassay methods. Reagents, devices, and kits using the anti-hapten antibodies are also disclosed. The present invention also relates to dyed erythrocytes, preferably fixed, which are coated with antibodies. Also disclosed is the use of these dyed erythrocytes in agglutination assays to detect and measure the presence of an analyte in a sample. The analyte can be a hapten, an antigen, or an antibody. Also included are agglutination assays, compositions and kits using these dyed and coated erythrocytes.

Abstract: Genotoxic chemicals are an existing wide-spread health hazard to the human population. Advances in genetic toxicology testing have made it possible to assay potential mutagens, carcinogens, teratogens and clastogens in the environment. The mouse micronucleus assay provides an example of an excellent test for genetic damage to cells. When chromosome breaks occur in the blood stem cell population, the damaged piece of chromosome remains behind as a micronucleus in the normally DNA deficient red blood cells. However, currently available manual micronucleus assays are costly, time consuming, and labor intensive. In addition, the statistics are often marginal since the number of micronucleii (MNs) in 1000 polychromatic cells are scored manually, yielding limited amounts of data. This invention discloses the means for assaying the change in micronucleated cells by high speed flow cytometry.

Abstract: A method for immunochemical determination of a hapten in a sample is disclosed, in which(A) a high-molecular compound to which the hapten is bound (reagent A),(B) insoluble carrier particles carrying thereon an antibody to the hapten (reagent B), and(C) magnetic substance-containing insoluble carrier particles carrying thereon an antibody to an antigenic determinant in the high-molecular compound and different from the hapten (reagent C),are used. These three reagents are dispersed in the sample, then, a magnetic field is applied to separate from the reaction mixture unreacted reagent (C) and agglutinated particles formed from the reagent (B) and the reagent (C) through the reagent (A). The amount of the reagent (B) remaining dispersed in the reaction mixture is measured, thereby determining the extent of competitive inhibition to the agglutination of the reagent (B) and the reagent (C) through the reagent (A) by the reaction between the hapten in the sample and the reagent (B).

Abstract: A method for drying mammalian cells, such as erythrocytes, lymphocytes, leukocytes and platelets onto a solid-phase support for use in solid-phase immunoassays through use of a drying solution and an article for use in immunoassays prepared by such method. The method comprises immobilizing a monolayer of cells onto the solid-phase support by non-covalent binding. This is accomplished by staining the solid-phase support with an organic dye having a net positive charge which permits non-covalent binding of the cells which carry a net negative charge to the solid-phase support. These cells are dried or fixed to the solid-phase support by addition of a drying solution which comprises an aqueous solution of a monosaccharide, disaccharide, trisaccharide or cyclitol and a salt. The preferred monosaccharide is D-(-)glucose and the preferred salt is sodium chloride. The preferred drying solution comprises a 1.0 M solution of dextrose and a 154 mM solution of sodium chloride.

Abstract: A reaction kit is provided with a reaction zone of suitable cross-sectional area to aspirate a sample by capillarity and a transparent plate with a flat surface in at least part of the reaction zone, and a supporting base to incline the reaction zone at a specified angle. The reaction zone is normally a space formed by two opposite transparent plates and spacers inserted between these two transparent plates. A supporting base may be molded separately, the transparent plates and spacers being set on the base at the time of measurement, or alternatively the base may be molded integrally with at least one of the plates defining the reaction zone. Further, substances with specificity such as antigens or antibodies may be coated on a surface in the reaction zone.

Abstract: The present invention describes a class of dyes for use in staining cell samples and methods of making such dyes. A preferred class of dyes known as detergent dyes which possess the ability to stain cells in whole blood and are only slowly leached or lost from the stained cells over time are described. The present invention has application, for example, to blood typing 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: The present invention relates to a method to detect feline blood type B comprising: combining feline blood with a sufficient amount of Triticum vulgaris lectin to agglutinate feline blood type B. Additionally, this invention relates to a kit to detect feline blood type B.

Abstract: Microcapsule-reagents are prepared by previously reacting at least a part of an antigen or antibody attached to microcapsules encapsulating a labeling substance with an antibody or antigen which is specifically reactive therewith, and then the reagent thus prepared is reacted with a sample containing an antigen or antibody in the presence of a complement, whereby highly sensitive immunoassay of the antigen or antibody in the sample can be realized even when the antigen or antibody attached to the microcapsules has a lowered activity or has only low reactivity with the antibody or antigen in the sample.

Abstract: A method for detecting and measuring the image of an agglutination particle pattern produced in a test liquid in which the existence of hemolysis in a test liquid is judged at the same time as an analysis is conducted of particle patterns produced in the test liquid in response to an immunological agglutination reaction. By this method, it is possible to increase the precision of the analysis of the particle pattern by feeding back the result of the judgement of the existence of hemolysis in the test liquid. A reaction apparatus suitable for use in the method according to the invention is also disclosed. In this apparatus, a plurality of wells for containing the test liquid are formed, and a plurality of marks are formed in positions where the marks can be read through the test liquid by an optical system for detecting the particle patterns produced in the test liquid. Therefore, hemolysis existing in the test liquid can be objectively judged.

Abstract: Quaternized derivatives of acridine orange and reagents incorporating such derivatives and their use in quantitatively determining reticulocyte levels in whole blood specimen by fluorescence flow cytometry techniques are disclosed. The quaternized derivatives of acridine orange are of the general formula: ##STR1## wherein Y is bromide (Br.sup.-) or iodide (I.sup.-), andX may be R.sub.1 and/or R.sub.2 substituted benzyl group ##STR2## R.sub.1 is hydrogen or fluorine, and R.sub.2 is fluorine, trifluoromethyl or hydrogen, or X may be hydroxyl ethylene.

Abstract: A method for determining the presence of a specific binding member bound to first particles in a liquid medium is disclosed. The method comprises providing in combination (1) a liquid medium suspected of containing a specific binding member bound to first particles, (2) means for agglutinating the first particles in relation to the presence of the specific binding member, and (3) second particles having the same or a different specific binding member for said means for agglutinating bound thereto, thereby providing for said means to agglutinate the second particles. Agglutination of the first and second particles are separately detectible and distinguishable by spectroscopic measurement. The medium is incubated and agglutination of each of the particles is determined spectroscopically without separating the first and second particles.

Abstract: Assay methods and compositions are provided for determining an analyte in a sample suspected of containing the analyte. The composition comprises in a novel single liquid reagent at least one specific binding pair (sbp) member and its complementary member wherein at least one sbp member is reversibly confined in a material that temporarily renders the confined sbp member incapable of binding with its complementary sbp member. At least one of the sbp members is bound to a member of a signal producing system capable of producing a detectable signal in relation to the amount of analyte in the sample. The confinement is reversed, any remaining members of the signal producing system are added, and the signal produced in relation to the amount of analyte is measured. Examples of the confining material are lipid bilayers, cells and gels.

Abstract: A reaction apparatus with sample inlet channel of suitable cross-sectional area for aspirating a sample into the interior of the apparatus by capillarity, a recess provided on the inner wall of the sample inlet channel, and a transparent plate with a flat surface arranged opposite the recess. The recess has sloping walls, and may for example, have a conical or hemispherical form. The recess may moreover be a V-shaped or U-shaped groove. Further, the inner wall of the recess may be coated with antibodies or antigens.

Abstract: System for quantifying components in a liquid sample, for example white blood cells in blood, based on use of substantially non-compressible beads of uniform diameter as spacers between a microscope slide and a cover slip. The distance between the slide and cover slip is determined by the diameter of the beads, and thus the volume of the sample being viewed can be precisely determined. Prior to use, the beads may be adhered to a microscope slide or stirring rod in a dried adhesive matrix which is soluble in the liquid sample to be examined.

Abstract: This invention relates to a flotation immunoassay employing a novel buoyant matrix to which an antigen or antibody is coupled and which separates the bound and free products of the assay by floating to the surface of the reaction liquid. The novel flotation device which makes it possible to detect and to quantitate either antigen or antibody can also be used to fractionate cells and molecules.

Abstract: A reagent for reticulocyte counting by flow cytometry which comprises two solutions, namely, a stock solution for staining in which a dye is dissolved in a nonaqueous solvent, and a buffer solution which satisfies the optimum staining conditions.By combining these two solutions immediately before measurement, a stable final staining solution for reticulocyte counting can always be obtained.

Abstract: A method for analyzing antigen or antibody contained in a sample by introducing a first end surface of at least one measuring light guide member and a second end surface of at least one reference light guide member in the sample. Each of said measuring and reference light guide members being coated with a protection layer except for said first and second end surfaces and an other end surface of respect light guides. Only on said first end surface of the measuring light guide member being fixed an antigen or antibody which is specifically reactive with the antigen or antibody contained in the sample to effect an antigen-antibody reaction. Photoelectrically detecting optical properties of said first and second end surfaces of the measuring and reference light guide members, respectively, by transmitting light through the meauring and reference light guide members to derive detection signals as a measure of antigen or antibody in the sample.

Abstract: A reagent for reticulocyte counting by flow cytometry characterized in that it contains a carbonate salt.If a blood sample is left exposed to ordinary air, non-specific staining of erythrocyte can be prevented with the addition of not less than about 1 mM of NaHCO.sub.3.

Abstract: An article for performing solid-phase immunological assays and method for performing assays utilizing the article of the invention. A solid-phase support suitable for immunological assays is stained with an organic dye that has a net-positive charge and a hydrophobic aromatic ring structure. An immunologically reactive component having a net-negative charge is immobilized by the dye to the support through non-covalent interactions. The immunologically reactive component is tightly bound by the organic dye. This is advantageous when performing immunological assays utilizing automated washing instruments. A sample of a biological fluid is added to the article having the immunologically reactive component bound thereto to detect the presence of antigens or antibodies in the biological fluid specific for the bound immunological component. The presence of the antigen-antibody complex may be determined by any known method.

Abstract: System for quantifying components in a liquid sample, for example white blood cells in blood, based on use of substantially non-compressible beads of uniform diameter as spacers between a microscope slide and cover slip. The distance between the slide and cover slip is determined by the diameter of the beads, and thus the volume of the sample being viewed can be precisely determined. Prior to use, the beads may be adhered to a microscope slide or stirring rod in a dried adhesive matrix which is soluble in the liquid sample to be examined.

Abstract: A method and apparatus for conducting specific binding pair assays, such as immunoassays, is described. A porous membrane capable of non-bibulous lateral flow is used as assay substrate; a member of the binding pair is affixed in an indicator zone defined in the substrate. The sample is applied at a position distant from the indicator zone and permitted to flow laterally through the zone; any analyte in the sample is complexed by the affixed specific binding member, and detected. A novel method of detection employs entrapment of observable particle in the complex. Blood is a particularly preferred sample as the red blood cells can be used as the observable particles for detection of the complex.

Abstract: A method of determining the presence and quantity of an analyte of interest in a particulate-containing sample is disclosed, as is a construct for use in the method. The method is particularly useful for determining an analyte in whole blood and in fermentation suspensions. The construct is comprised of a first moiety, which is a particulate-binding moiety and a second moiety, which binds the analyte of interest.