Abstract: A method for protein determination utilizing a pyrocatechol violet-metal complex, a composition used in the protein determination assay, and a kit.

Abstract: Assay methods are disclosed involving specific binding reactions which are simplified compared to known methods. A compound capable of producing chemiluminescence is immobilized on a solid support as is a member of a specific binding pair for capturing an analyte from a sample. An activator compound that activates the chemiluminescent compound and is conjugated to a specific binding pair member is added in excess along with the sample to the solid support. Addition of a trigger solution causes a chemiluminescent reaction at the sites where the activator conjugate has been specifically bound. The assay methods are termed non-separation assays because they do not require removal or separation of excess detection label (activator conjugate) prior to the detection step. The methods are applicable to various types of assays including immunoassays, receptor-ligand assays and nucleic acid hybridization assays.

Abstract: The invention describes biomarkers which can be used to predict the likelihood that an individual will develop Diabetes. The biomarkers can also be used to screen large groups in order to identify individuals at risk of developing Diabetes.

Abstract: The present invention relates to a method of extracting tacrolimus in a test blood sample to use in a tacrolimus quantification assay. The method of the present invention extracts tacrolimus in non-precipitating, non-denaturing aqueous environment, avoiding cumbersome manual pretreatment procedures or the use of organic solvents. The method of the present invention permits full automation of tacrolimus quantification assays.

Abstract: A multifunctional reagent for erythrocytes containing an amount sufficient to produce the lysis of erythrocytes or the sphering of erythrocytes in such a way that they can be detected by a cytometer or an automatic counting device, of a carbamate or of an agent inducing the formation by the erythrocytes, from carbonate and from a nitrogenated heterocycle or ammonium ions, of a carbamate combined with the absorption of CO2 by the erythrocytes, process for lysing or sphering erythrocytes and preparation process for leucocytes.

Abstract: The invention relates to a reagent and a process for the identification and counting of biological cells in a sample. This reagent comprises a cell lysing agent selected from at least one detergent in a concentration capable of specifically lysing a given type of cells in the sample, and a stain capable of marking the intracellular nucleic acids of the remaining unlysed cells. Application in particular for the identification and counting of cells using an automated analysis system based on flow cytometry.

Abstract: An automated in situ heat induced antigen recovery and staining method and apparatus for treating a plurality of microscope slides. The process of heat induced antigen recovery and the process of staining the biological sample on the microscope slide are conducted in the same apparatus, wherein the microscope slides do not need to be physically removed from one apparatus to another. Each treatment step occurs within the same reaction compartment. The reaction conditions of each reaction compartment for treating a slide can preferably be controlled independently, including the individualized application of reagents to each slide and the individualized treatment of each slide. The reagents are preferably held in a reagent dispensing strip similar to a “blister pack”.

Abstract: An automated in situ heat induced antigen recovery and staining method and apparatus for treating a plurality of microscope slides. The process of heat induced antigen recovery and the process of staining the biological sample on the microscope slide are conducted in the same apparatus, wherein the microscope slides do not need to be physically removed from one apparatus to another. Each treatment step occurs within the same reaction compartment. The reaction conditions of each reaction compartment for treating a slide can preferably be controlled independently, including the individualized application of reagents to each slide and the individualized treatment of each slide. The reagents are preferably held in a reagent dispensing strip similar to a “blister pack”.

Abstract: The present invention provides methods, diagnostic assays, and diagnostic kits based on said methods, to determine levels of immunosuppressive complexes containing immunosuppressive drugs tacrolimus, sirolimus and cyclosporine A separately and in combination, formed in the blood of a drug-treated patient or in a patient candidate to immunosuppressive drug therapy. These methods, assays and kits are especially useful when using automated systems.

Abstract: The present invention relates to a method of extracting tacrolimus in a test blood sample to use in a tacrolimus quantification assay. The method of the present invention extracts tacrolimus in non-precipitating, non-denaturing aqueous environment, avoiding cumbersome manual pretreatment procedures or the use of organic solvents. The method of the present invention permits full automation of tacrolimus quantification assays.

Abstract: The present invention provides an assay, suitable for neonatal screening of hemoglobinopathies by quantitatively determining the presence of at least one pair of hemoglobin (Hb) variants, determining the ratio between the signals obtained from each variant; and, based on that ratio, determining whether the tested subject is afflicted or not.

Abstract: New applications for the use of distinguishable particulate labels available in a variety of hues and sized in the submicron range are described. These applications include profiling of cellular components, obtaining secretion patterns, identifying a multiplicity of components in chromatographic or electrophoretic techniques and identification of desired immunoglobulin secreting cells.

Abstract: A B-lymphocyte classifying method includes the steps of (1) preparing a measurement sample by mixing a blood sample with a lysing reagent to lyse erythrocytes and to shrink the B-lymphocytes in the blood sample, (2) letting the measurement sample flow through a flow cell of a flow cytometer, (3) radiating a light beam onto a cell in the measurement sample that is flowing through the flow cell, (4) detecting at least two scattered light emitted from the irradiated cell, (5) specifying a region of a B-lymphocyte cluster in accordance with the detected scattered light, and (6) counting the number of B-lymphocytes in the specified region.

Abstract: A method and apparatus for preparing biological cell samples for intracellular analysis. The invention is based upon the recognition that many of the steps of the conventional methods for such sample preparation can be eliminated, leading to a process that readily lends itself to automation and the advantages associated therewith. The method of the invention comprises the steps of (a) cell-fixation, (b) permeabilization and (c) staining (or labeling) of intracellular molecules of interest by probes that are readily detectable by flow cytometric techniques, all without any intervening cell-washing (and re-suspension) steps. Rather, the single cell-washing step is effected after these three steps have been carried out. Preferably, the washing step is carried out by passing the fixed, permeabilized and stained cell sample through a semi-permeable membrane that serves to filter out (by transmission) interferants to waste while retaining the cells of interest.

Abstract: A reagent for partially lysing a cell membrane of a red blood cell infected with a malaria parasite, comprising: a first surfactant which has predetermined lysing ability for the red blood cell membrane; and a second surfactant which has weaker lysing ability than that of the first surfactant; wherein the reagent has pH of 5 to 7 and osmotic pressure of 200 to 300 mOsm/kg·H2O; and wherein the reagent partially lyses the malaria infected red blood cell membrane such that a malaria parasite is retained in the red blood cell and fluorescent dye passes through the cell membrane, is disclosed. A reagent for detecting malaria infected red blood cells, and a sample analyzing method for detecting malaria infected red blood cells, are also disclosed.

Abstract: The invention provides a flow cytometric method for measuring dendritic cell function in whole blood, comprising: (a) contacting a whole blood sample with a dendritic cell activator; (b) adding to the sample a plurality of dendritic cell-distinguishing antibodies and at least one cytokine-specific antibody; and then (c) flow cytometrically assaying the sample for the binding of the cytokine-specific antibody by at least one distinguishable DC subset. Other assays are presented in which DC activation markers are assessed.

Abstract: In a method for measuring an analyte, which comprises a reaction step of forming a reaction system including a sample containing whole blood, a first substance carried by a solid carrier and specifically binding to an analyte contained in the sample and a second substance specifically binding to the analyte and allowing the analyte to react with the first and second substances and a measurement step of measuring the formed reaction product, (1) the reaction step is performed in a state that blood cells are not disrupted, and (2) at least the reaction step is performed in the presence of a sufficient amount of a detergent that does not cause hemolysis, does not inhibit reactions of the analyte with the first and second substances specifically binding to the analyte and can prevent influence on the reaction system of a component existing in the reaction system.

Abstract: This invention relates to an improved procedure for the multiparameter analysis of leukocyte subpopulations from peripheral blood, bone marrow or any body fluid containing blood nucleated cells, including the quantification of each subpopulation in terms of cells per microliter. The method uses a cocktail of at least four monoclonal antibodies labelled each with a different fluorescent tag, the measurement of at least two light scatter and four fluorescence emissions related parameters and at least one population of fluorescent beads to differentiate and enumerate between and among the different leukocyte subpopulations in the peripheral blood, the bone marrow or other body fluid.

Abstract: A new method of cell fixation using a mild denaturing heat and the fixative PERMIFLOW™ or similar fixative that preserves cell morphology, light scatter profiles, nucleic acid content, cell surface epitopes and unmasks internal epitopes previously not available. The method can be combined with analysis of each of these parameters by flow cytometry and thus has application in drug screening and patient or tumor monitoring protocols.

Abstract: A system for detecting a particle disposed in a detection area. The system includes a light-emitting source for generating light. The light is directed at the particle. The system further includes a modulator configured to in-situ modulate at least one environmental parameter of the particle to alter a detectable response of the particle. The modulator provides an enhancement in detection selectivity of the particle in the presence of interfering particles and species. Further, the system includes a detector configured to detect alteration in the detectable response of the particle.

Abstract: The embodiments of the invention are directed to a SERS cluster comprising a capture particle that is at least partially surrounded by analyte molecules, wherein both the capture particle and the analyte molecules surrounding the capture particle are at least partially surrounded by enhancer particles, wherein a majority of the analyte molecules are either sandwiched between capture and enhancer particles or located between junctions of the enhancer particles. The embodiments of the invention also relate to methods of manufacturing and detecting the SERS cluster. The embodiments of the invention also relate to a SERS active particle comprising a tag molecule comprising a Raman active compound and a probe or a linker having a specific biochemical binding capability and to a method for detecting of a target molecule using a SERS active particle having a tag molecule comprising a Raman active compound and a probe or a linker.

Abstract: A method is provided for determining the level of an analyte in the blood of an individual based on determination of the level of the same analyte in non-blood sample (e.g. urine, saliva and hair) obtained from the individual. The non-blood sample contains red blood cells and the volume of the blood in the sample together with the amount of the analyte in the sample are the basis for calculating the level of the analyte in the individual's blood. Kits for carrying out the above method are also provided.

Abstract: The invention concerns a reagent for determination of leucocytes and measurement of haemoglobin in a sample of blood. This reagent comprises a buffer system that is suited to adjust selectively the pH of the reagent to an acidic value, at least one detergent of cationic type, a nitrogenous compound and, optionally, at least one inorganic salt. This reagent can be used in haematological analyses in human medicine and also permits the identification of a leucocytic subpopulation, in particular the basophil polymorphonuclear leucocytes.

Abstract: A whole blood immunoassay includes the steps of mixing a whole blood sample with sensitized insoluble carrier particles to cause an immune agglutination; diluting the resulting agglutination mixture with an aqueous solution containing an erythrocyte lysing agent to lyse erythrocytes, thereby preparing an assay sample; and determining a degree of agglutination of the assay sample.

Abstract: The present invention recognizes that separation of components of a sample facilitate, and are often necessary for, sample analysis. Dielectrophoretic separation provides an efficient, reliable, nondisruptive, and automatable method for the separation of moieties in a sample based on their dielectric properties. The present invention provides compositions and methods for enhancing the dielectrophoretic separation of one or more moieties in a sample. A first aspect of the present invention is a solution that when mixed with a sample, modifies at least one dielectric property of one or more components of the sample and has a conductivity such that one or more moieties of the sample can be separated using dielectrophoresis. Such solutions can be used in the analysis of samples on chips, and can be used in methods that use binding partners, including microparticles that can be translocated by dielectrophoretic forces, traveling-wave dielectrophoretic forces or magnetic forces.

Abstract: A method for counting leukocytes which comprises liberating elastase from granulocytes contained in a specimen, adding an anti-granulocyte elastase antibody to the thus liberated elastase, measuring the antibody bonded to the elastase to thereby determine the concentration of the elastase, and then calculating therefrom the number of leukocytes contained in the specimen with the use of the ratio of the leukocyte count to the elastase concentration. This method makes it possible to conveniently and less expensively count leukocytes at a high accuracy comparable to the one established by using a conventional automatic blood cell counter, as the figure shows.

Abstract: Disclosed are materials and methods for detecting biomolecules in samples employing transponders associated with the bead(s) used as the solid phase in the assay, and information pertinent to the assay is encoded on the transponders memory elements. A dedicated read/write device is used remotely to encode or remotely to read the information. 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 process for permeabilizing erythrocytes in which the erythrocytes are subjected successively to the action of (a) a fixing agent containing an aliphatic aldehyde and oligosaccharide, (b) a permeabilizing agent containing a detergent and an oligosaccharide, kit for permeabilizing erythrocytes, kit for immuno-marking fetal erythrocytes, and a process for identifying fetal erythrocytes by immuno-marking.

Abstract: An automated in situ heat induced antigen recovery and staining method and apparatus for treating a plurality of microscope slides. The process of heat induced antigen recovery and the process of staining the biological sample on the microscope slide are conducted in the same apparatus, wherein the microscope slides do not need to by physically removed from one apparatus to another. Each treatment step occurs within the same reaction compartment. The reaction conditions of each reaction compartment for treating a slide can preferably be controlled independently, including the individualized application of reagents to each slide and the individualized treatment of each slide. The reagents are preferably held in a reagent dispensing strip similar to a “blister pack”.

Abstract: The invention features a novel method of assaying glycated hemoglobin (GHb) in a sample using a capture molecule that binds GHb and hemoglobin (Hb), and detecting bound GHb with a molecule that distinguishes GHb from non-glycated hemoglobin (Hb). The molecule used to distinguish GHb from Hb may be an antibody. The assay and antibodies of the invention are useful for the evaluation of GHb in disease states such as diabetes.

Abstract: A compact combination blood cell count and immunoassay apparatus for measuring drawn whole blood specimen includes a load station for receiving the whole blood specimen and a nozzle member that can selectively withdraw whole blood from the blood specimen. A blood cell count section and an immunoassay section are mounted along a unitary path of movement of the nozzle member so that a single nozzle member may be used for transporting the whole blood to the respective blood cell count section and immunoassay section. A controller can automatically direct a transport mechanism to use only the whole blood and to perform various steps of adding dilution liquid and reagents, along with appropriate washing of the nozzle and sample cells to insure an automatic operation. A hematozorit value can be determined from the blood cell count signal and used to correct the immunoassay signal when whole blood is used.

Abstract: A lysis chamber of an assay device capable of producing lysis of cells in a sample fluid such as whole blood, said chamber comprising a surface which contacts the sample fluid when the sample fluid is placed into the assay device; and a lytic material immobilized on the surface, whereby cells of the sample fluid are lysed when they contact the lytic material. The chamber can delimit a capillary space, and lytic material can be saponin or a detergent. Methods employing devices comprising such chambers can assay for whole blood amounts of cyclosporin or hemoglobin A1c.

Abstract: A device for analyzing a whole blood sample is provided. The device comprises a conventionl hematology analyzer integrated with a fluorescence cyometry analyzer. A controller is provided for controlling the analyzers, obtaining and utilizing data from both and reporting a quantitative result. Methods are also provided for analyzing a whole blood sample. One such method comprises the steps of performing on a single instrument an analysis of impedance associated with the blood sample, an analysis of light scatter associated with the blood sample, and an analysis of fluorescence associated with the blood sample. Data is collected and utilized. A result is reported.

Abstract: Provided are automated methods for distinguishing and differentiating cells in a whole blood sample. In one of the methods, a whole blood sample is provided. One or more tests to be performed on the whole blood sample is selected. The tests to be performed on the whole blood sample are correlated. A volume of the whole blood sample is aspirated into an automated instrument system which automatically performs conventional hematology analysis and fluorescent cytometry analysis on the whole blood sample. A first aliquot of the whole blood sample is dispensed into at least one sample receiving vessel. The first aliquot of the whole blood sample is mixed with a fluorescent reagent. The first aliquot of the whole blood sample mixed with fluorescent reagent is diluted and transported through a flow transducer system.

Abstract: A method for immunologically determining antistreptolysin O antibody comprises contacting a test sample solution containing antistreptolysin O antibody with a binding agent to selectively bind the antistreptolysin O antibody to the binding agent, and determining an amount of the bound antistreptolysin O antibody, wherein the binding agent comprises a streptolysin O-immobilized carrier obtained by contacting a solution containing streptolysin O with a carrier having immobilized thereon at least one steroid represented by formula (1): ##STR1## wherein R.sup.1 is a side chain moiety of cholesterol or a side chain moiety of cholic acid, wherein each of the side chain moieties is independently unsubstituted or substituted and independently saturated or unsaturated; R.sup.2 is hydrogen or hydroxyl; and each dashed line independently represents a single bond or no bond.

Abstract: The invention provides compounds which specifically inhibit factor Xa activity. The compounds consist of the structure X.sub.1 -YIR-X.sub.2, wherein X.sub.1 is H, acyl, alkyl, acylalkyl, arylalkyl or one or more amino acids, and X.sub.2 is a modified C-terminal group, one or more carboxy-protecting groups or one or more amino acids or other substituent, and Y, I and R are tyrosine, isoleucine and arginine, respectively, or peptidomimetic or organic structures that possess the same functional activity as Y, I and R, respectively. In addition, the present invention provides a compound having the structure A1-A2-(A3).sub.m --B, where m is 0 or 1. A compound of the invention can be linear or cyclic and can be about 2 and 43 residues in length. A compound of the invention is characterized, in part, in that it exhibits a specific inhibition of factor Xa activity with a K.sub.i of .ltoreq.100 .mu.M, preferably .ltoreq.

Abstract: A solid phase method of detection or assay of the presence or amount in a serum or plasma sample of a target antibody specific to a cell surface antigen. The sample is contacted with an immobilised preparation of cells bearing the antigen and antibody bound thereto is detected or assayed by means of an indicator comprising a binding partner for the antibody bound to labelled latex particles.

Abstract: The invention is a rapid, continuous test for glycated hemoglobin using a non-equilibrium affinity binding method. Agarose beads derivatized with 3-aminophenylboronic acid specifically bind glycated hemoglobin. This solid phase is incorporated into a sample processor card, modified to mix and to separate the test solution from the solid phase prior to absorbance readings. Two absorbance readings are made on the test solution, one immediately after mixing the reagent/diluent with the specimen, and one after a significant amount of binding has occurred. A linear correlation between total glycated hemoglobin and hemoglobin A.sub.1c permits standardization and reporting of units equivalent to % hemoglobin A.sub.1c. Stable glycated hemoglobin solutions for use as standards in the assay, and a method for preparing the standards are also disclosed.

Abstract: A device for analyzing a whole blood sample is provided. The device comprises a conventionl hematology analyzer integrated with a fluorescence cyometry analyzer. A controller is provided for controlling the analyzers, obtaining and utilizing data from both and reporting a quantitative result. Methods are also provided for analyzing a whole blood sample. One such method comprises the steps of performing on a single instrument an analysis of impedance associated with the blood sample, an analysis of light scatter associated with the blood sample, and an analysis of fluorescence associated with the blood sample. Data is collected and utilized. A result is reported.

Abstract: This invention provides a method for preserving cells which comprises the steps of (a) suspending cells in a physiologically-acceptable, isotonic medium; and (b) fixing the cells so suspended at a temperature of less than about 10.degree. C. under sufficiently hypertonic conditions so as to disperse the cells in a single, unagglutinated state, thereby preserving cells. This invention also provides a method for detecting cells separated from a sample which have been preserved according to the aforementioned method. This invention also provides a method for visualizing cells. Also provided is a method for detecting a metabolic process in cells present in a sample. This invention also provides a method for detecting the presence of rare cells in a sample which specifically possess on their surfaces a moiety recognized by a known ligand comprising preserving cells separated from the sample according to the aforementioned method for preserving cells.

Abstract: A device for analyzing a whole blood sample is provided. The device comprises a conventional hematology analyzer integrated with a fluorescence cytometry analyzer. A controller is provided for controlling the analyzers, obtaining and utilizing data from both and reporting a quantitative result. Methods are also provided for analyzing a whole blood sample. One such method comprises the steps of performing on a single instrument an analysis of impedance associated with the blood sample, an analysis of light scatter associated with the blood sample, and an analysis of fluorescence associated with the blood sample. Data is collected and utilized. A result is reported.

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: A hematology control product comprising leukocyte analogs and an aqueous solution of a plasma substance for use in a blood counting instrument is described. The instrument employs a lytic reagent system for the lysable red blood cells in the control product. The plasma substance is in an amount effective to enable the differentiation of each of said leukocyte analogs relative to the physical attributes of the analogs, said physical attributes of the analog are similar to human leukocytes. Preferably, the plasma substance comprises cholesterol or its derivatives.A method for using the cell suspension media comprising an aqueous solution of a plasma substance is also described. The method provides a quality control to determine whether an instrument is operating within manufacturer's specifications.

Abstract: A non-toxic and non-cytolytic derivative of streptolysin O which retains at least one (preferably immunodominant) epitope is produced, and can be used especially in diagnostic tests for detecting of the presence of antibodies to Streptococcus pyogenes in a sample.

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 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: 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 method for detecting pathogen infection in a host is provided. The method comprises lysing phagocytes from the host to release soluble components of the pathogen which are detected subsequently using a specific binding assay.

Abstract: An analytical method for determining the relative amount of a particular hemoglobin derivative, e.g., glycated hemoglobin, in a blood sample wherein the amounts of both total hemoglobin and the hemoglobin derivative are measured and related mathematically, e.g., as a percentage. The blood sample is treated with the combination of a thiocyanate salt and an oxidant to denature the hemoglobin in the sample and to convert hemoglobin to met-hemoglobin. Met-hemoglobin is measured spectrophotometrically to give the amount of total hemoglobin present, and the denatured hemoglobin derivative can be distinguished and measured by immunoassay. The presence of the oxidant in the denaturant solution has been found to increase the rate of enaturation of hemoglobin thereby reducing the overall assay time and improving the reliability of the determination.

Abstract: A lysing agent and a method for utilizing the lysing agent in the identification and enumeration of cells of a select subclass of leucocytes is provided. The lysing agent includes formaldehyde, an alkali or alkaline earth salt of a weak acid and a polyhydric alcohol.