Abstract: A method for determining normal intrauterine pregnancy during the first 20 weeks of pregnancy comprises obtaining a test sample; and determining the presence of a fetal restricted antigen in the sample. The test sample is removed the vaginal cavity in the vicinity of the cervical canal and/or the cervical os. One fetal restricted antigen is fetal fibronectin.In one embodiment of this invention, the test sample is contacted with an insoluble support to which anti-(fetal restricted antigen) antibody is adhered, and the fetal restricted antigen binding to the support is determined. Alternatively, the test sample is contacted with an insoluble support to which is adhered an antibody which binds a class of substances including the fetal restricted antigen; and the fetal restricted antigen binding to the support is determined. Reagents and reagent kits are also included.

Abstract: A test kit useful for the determination of an immunological ligand includes a labeled receptor for that ligand, a specific wash solution and a test device. The wash solution is buffered to a pH of from about 5 to about 9 and contains at least 0.01 weight percent of a compound having a dodecyl sulfate anion and an alkali metal or ammonium cation. One such compound is sodium dodecyl sulfate. The test device has several test wells, and in each well there is a microporous membrane. A receptor for the ligand of interest is immobilized in one test well. The kit is particularly useful for measuring human chorionic gonadotropin as an early indicator of pregnancy.

Abstract: A buffered aqueous composition is useful simultaneously as a wash solution and a dye-providing composition in specific binding assays involving enzyme-labeled specific binding reagents. The wash composition includes a dye-providing composition, a buffer and an organic solvent having a certain molecular weight and water-solubility. Another useful composition includes a particulate substrate having avidin attached thereto, and a peroxidase reducing agent. Either composition can be provided in a diagnostic test kit, and can be used to detect a specific binding ligand in assays.

Abstract: A device to determine human chorionic gonadotropin (hCG) in urine is described.

Abstract: A substance having an inhibitory effect on the production of human choriogonadotropin (hCG) is described, which comprises a protein isolated from human decidual cells with a molecular weight of about 7,000 to about 10,000 daltons, and useful for use in inhibiting production of hCG in vivo or in vitro or may be measured as an indication of a cause of hCG inhibition.

Abstract: A general method of assay for biological molecules using daylight fluorescent particles. The method described is applicable to assays involving immunological reagents, nucleic acids, hormones and neurotransmitters.

Abstract: The present invention describes methods and compositions for detecting the fertility of a canine female based on identification of certain characteristic cLH peptides and carrier proteins found in canine urine. More specifically free or complexed metabolites of luteinizing hormone (LH) are detected.

Abstract: A method for determining pregnancy in a dog or cat, as well as a method for distinguishing between pseudopregnancy and actual pregnancy in a dog or cat which methods comprise measuring relaxin levels in bodily fluids or tissues which carry relaxin early in pregnancy, the presence of significant amounts of relaxin being indicative of pregnancy.

Abstract: The invention disclosed herein relates to the field of one-step assays and latex agglutination tests ("LAT"), and provides an improved method of preparing and using a one-step immunoassay, as well as improved methods of preparing coated latex particles for use in diagnostic assays, and especially, immunochromatographic assays.

Abstract: A method for determining increased risk of labor and fetal membrane rupture after week 20 of pregnancy comprises obtaining a secretion sample from the vaginal cavity; and determining the presence of a fetal restricted antigen in the sample. The sample can be removed from anywhere in the vaginal cavity, but is preferably removed from the posterior fornix or and/or cervical os. One fetal restricted antigen is fetal fibronectin. In one embodiment of this invention, the sample is contacted with an insoluble support to which anti-(fetal restricted antigen) antibody is adhered, and the fetal restricted antigen binding to the support is determined. Alternatively, the class of substances of which the fetal restricted antigen is a member is captured with a general binding antibody (such as anti-human fibronectin antibody), anti-(fetal restricted antigen) antibody (such as anti-fetal fibronectin antibody) is conjugated with the support, and binding with fetal restricted antigen is determined.

Abstract: The invention provides a process and products for the detection of the C peptide of relaxin in the urine of animals, as a positive indication of pregnancy. The invention may employ monoclonal antibodies generated to various epitopes on the C peptide.

Abstract: Preeclampisa, pregnancy induced hypertension (PIH) and eclampsia are determined by identifying the presence of an endothelial cell marker in a sample of blood, plasma or serum of a pregnant woman using a sandwich or competition immunoassay. Cellular fibronectin marker in a sample is determined by binding with an anti-(cellular fibronectin) antibody. Reagents for these methods are also an aspect of the invention.

Abstract: A general method of assay for biological molecules using daylight fluorescent particles. The method described is applicable to assays involving immunological reagents, nucleic acids, hormones and neurotransmitters.

Abstract: Provided is a test kit for conducting a membrane sandwich immunoassay of a multivalent antigen, optionally including reagents, the test kit includes a hollow container for reception of a fluid specimen, containing in series fluid absorbent body, a fluid pervious layer, and a fluid pervious solid membrane layer on which antibody is immobilized such that fluid added (e.g., labeled antibody and fluid specimen) to the container migrates through the membrane and in the presence of unlabeled scavenger antibody on the reactive surface of the membrane forms a multiple-site immunospecific antibody-antigen-antibody sandwich immobilized and labeled thereon, preferably in the form of a visible positive or negative indicator of the presence and/or amount of the analyte antigen in the specimen sample. The scavenger antibody serves to prevent false positive or other spurious reactions due to antigen analogs, antigen interference substances, or circulating human immnoglobulins.

Abstract: Monoclonal receptors raised to immunogenic polypeptides whose amino acid residue sequences correspond to sequences of oncoprotein ligands are disclosed, as are method for the production of those receptors and products and methods that utilize them. The monoclonal receptors bind both to the oncoprotein ligand to a portion of which the polypeptide corresponds in sequence, and to the immunogenic polypeptide to which the receptors were raised.

Abstract: The invention relates to an immunometric assay for a multivalent antigen in a sample which comprises forming a complex of the antigen together with multiple immobilized monoclonal antibodies against different epitopes of the antigen and with a detectably labeled soluble monoclonal antibody which is identical to one of the multiple immobilized antibodies. The labeled antibody associated with the complex is separated from the remaining soluble antibody and the detectably labeled antibody associated with the complex or unassociated with the complex is detected. Any one of the multiple immobilized monoclonal antibodies shows, by itself, substantially less binding towards the antigen in the immunometric assay, when used with itself or another monocolonal antibody in soluble labeled form, than when used with the multiple immobilized antibodies in combination.

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

Abstract: The present invention relates to a method of diagnosis of pathological pregnancy which relies on an evaluation of the amount of placental isoferritin (PLF) in the serum or amniotic fluid of a pregnant woman. Diagnosis may also be achieved by observation of percentages of PLF-bearing lymphocytes in the pregnant female. Detection of PLF levels is achieved by immunoassay with a PLF-specific anitbody. Also described is a method of treating or preventing pathological pregnancies and transplant or graft rejection by administration of effective amounts of PLF and/or a PLF-specific antibody in combination with immunization.

Abstract: A method for the determination of a free protein subunit of a quaternary protein in a sample, which comprises:(a) contacting a sample with a first immunological binding partner which is or will be bound to a carrier, wherein the first immunological binding partner binds epitopic determinants bindable only on the free protein subunit;(b) incubating the components of step (a) for a period of time and under conditions sufficient to form an immune complex between the free protein subunit, the first immunological binding partner, and the carrier;(c) separating the carrier of step (b) from the sample;(d) adding to the carrier of step (c), a detectably-labeled second immunological binding partner, wherein the second immunological binding partner binds epitopic determinants bindable on both the free protein subunit and the quaternary protein; and(e) determining the detectably-labeled second immunological binding partner in the carrier or in liquid phase.

Abstract: Detection of bindable substances such as antibodies and antigens using enzyme linked immunosorbent assays having particular utility in home diagnostic applications. The preferred implementation of the invention is characterized by the steps of admixing a sample suspected of containing the bindable substance to be detected with an antibody-enzyme conjugate, immersing an antibody coated solid support into the mixture and then exposing the coated support to an activated chromogenic solution. The conjugate for use in the home diagnostic assay is preferably contained within a lyophilized mixture. The lyophilized mixture contains components which preserve the antibody-conjugate's reactivity and immunologic binding specificity even if the lyophilized mixture had been subjected to hot, humid environmental conditions. Active components in the lyophilized mixture include polyethylene glycol, sugars, and surfactant.

Abstract: An isolated, substantially pure bovine pregnancy antigen for detecting and determining pregnancy in cattle consisting of a glycoprotein obtained from a pregnant bovine animal which is characterized by having a specific immunological reaction with a monoclonal antibody directed specifically against said glycoprotein wherein said monoclonal antibody is produced from hybridoma ATCC HB 8846.

Abstract: A method is presented for detecting placental dysfunction that is diagnostic of aneuploid chromsomal abnormalities. It is particularly useful for diagnosing pregnancies at risk for fetal aneuploid chromosome abnormalities, and consists of quantitating the hormone human chroionic gonadotropin (HCG) alone or in combination with the free alpha subunit of HCG (alpha-HCG). In a preferred embodiment of the invention, bodily fluids from women between 18 and 25 weeks of gestation are assayed using an immunoassay. Levels of 2.5 or more multiples of the median value for normal pregnancies (MOM) for HCG and/or alpha-HCG are indicative of fetal chromosome abnormalities. This test will detect approximately 70% of fetuses with chromosome aneuploidy.

Abstract: An immunoassay for detecting and measuring hCG in a sample includes an antibody directed to the carboxy terminal portion of the .beta. subunit of hCG and a monoclonal antibody directed to a determinant on hCG at a locus sufficiently remote from the carboxy terminal portion of the .beta. subunit of hCG that both antibodies can simultaneously bind to hCG, wherein at least one of the antibodies is delectable when both are bound to hCG.In a presently preferred embodiment, an immunoassay for hCG or hCB.beta. in urine includes a purified, labeled or detectable serum-derived antibody directed to the carboxy-terminal portion of the .beta. subunit of hCG and a matrix-bound monoclonal antibody directed to a locus on the .beta. subunit sufficiently remote from the carboxy-terminal portion that both antibodies can simultaneously bind to hCG or hCG.beta..

Abstract: A test kit is used in an agglutination immunoassay to determine a multivalent immune species, such as Streptococcus A antigen, in a biological sample. The method includes contacting an aqueous solution of the species with an agglutination indicator reagent having receptor molecules reactive with the species to form an agglutinate of the reaction product of species and receptor. These receptor molecules are bound to polymeric particles which contain tracer molecules. The resulting agglutinate is captured on a microporous membrane which has an average pore size which is at least five times greater than the average diameter of the polymeric particles. Unagglutinated residual materials are washed through the membrane using a wash solution. This solution has a pH of from about 5 to about 10 and an ionic strength of at least about 0.25. Tracer is then determined either in the agglutinate or in the residual materials.

Abstract: A highly sensitive and specific monoclonal-immuno-radiometric assay (M-IRMA) for hCG, using monoclonal antibodies (Mabs) directed against a 37-amino acid synthetic polypeptide analogous to the carboxyl terminus (CTP) of beta-hCG. Accordingly, in one embodiment, a method is described for the determination of human chorionic gonadotrThe present invention was made utilizing funds of the United States Government. The U.S. government is therefore granted a royalty-free, non-exclusive, world wide, paid-up license in this invention.

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

Abstract: A bovine pregnancy antigen, determined to be a glycoprotein, has been isolated and purified. If is diagnostic for the presence of pregnancy in cattle when detected by the use of antibodies to the antigen.

Abstract: There is described an enzyme inhibitor labelled immunoassay for measuring the concentration of an analyte in a sample wherein the substrate for the enzyme forms at least a part of the sample. In a particular embodiment the sample comprises or consists of a milk sample and the inhibitor label is capable of inhibiting the activity of an enzyme capable of clotting milk. Examples are given of suitable inhibitors. The assay described may be used to measure the concentration of progestogens or oestrogens in milk using the techniques of heterogeneous or homogeneous enzyme inhibitor labelled immunoassay. The results of such an assay give an indication of the fertility of a milk producing domestic animal (e.g. a cow) and may be used to diagnose pregnancy of such an animal. Particular compounds for use in the assay are described, as is a kit of reagents for use in the assay.

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: Compounds useful in a simultaneous multiple assay for analytes such as steroids, proteins, peptides, carbohydrates or drugs. The compound or compounds are prepared by labelling an individual analyte with a radioisotope through a chelating agent to form a coordinated compound. The assay uses one or more chelated labelled analytes with one or more labelled analytes wherein each radioisotope is different.

Abstract: A method and device for detecting an antigenic material in which the device comprises a test utensil having an indentation in which two reagent spots are placed, the first body being a dyed substrate having a coating of an antibody or antibody-like material thereon and the second of the two reagent spots comprising a dyed test-inert material or a dyed substrate with a coating of a normal animal serum, the dye employed in the second reagent spot having a different color than that employed in the first spot. When a liquid test sample is added to the indentation, the dyed substrate particles or components are suspended or solubilized, and the resulting suspension gives the appearance of a third color. A positive agglutination test is indicated by the formation of at least one spot having the color of the first dyed substrate against a background having the color of the second dyed substrate.

Abstract: A latex agglutination procedure for detecting and determining one of the participants in an antigen-antibody reaction, in which non-specific reactions are suppressed by additives, and an agent for carrying out this type of procedure are described.

Abstract: There is described an enzyme immunoassay for measuring the concentration of an analyte in a sample wherein the substrate for the enzyme forms at least a part of the sample. In a particular embodiment the sample comprises or consists of a milk sample and the enzyme is an enzyme capable of clotting milk. An example given of such an enzyme is chymosin. The assay described may be used to measure the concentration of progestagens or oestrogens in milk using the techniques of heterogeneous or homogeneous enzyme immunoassay. The results of such an assay give an indication of the fertility of a milk producing domestic animal (e.g. a cow) and may be used to diagnose pregnancy of such an animal. Particular compounds for use in the assay are described, as is a kit of reagents for use in the assay.

Abstract: The present invention relates to a method which eliminates centrifugation and decantation steps, to be performed in an automatic manner for carrying out specific binding assay tests, wherein liquid and solid phases are present.According to the invention, use is made of a specially designed device, consisting of a mixing reservoir into which is fitted snugly a mixer separator having a channel in the vertical axis of the mixer-separator. A rack holding a number of said mixing reservoirs containing the incubated reagents and analytes, capped with the mixer separators, is placed into a press-device designed to perform at a controlled rate a downward movement. The mixer separators are pushed downwards into the mixing reservoirs at a chosen rate for a preselected distance to complete the mass transport and separation operations. The separation devices are removed and either one of the separated phases can be measured in the desired analytical instrument for a quantitative or qualitative determination.

Abstract: In a particle agglutination assay for an antigen (Ag), there is included in the mixture a limited amount of a substance which binds univalently with a proportion of the Ag present, that Ag which is so bound being unable then to cause agglutination of the particles. In this way, unusually large concentrations of Ag can be assayed in that a proportion of the Ag is bound to the univalent substance and the particle agglutination assay is in effect conducted on the smaller amount of Ag still remaining free in solution.

Abstract: Early pregnancy can be determined by detection of a protein in serum, 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: A method and reagent system for detecting pregnancy in a mammal at an early stage by detecting enhanced activation of blood platelets.

Abstract: In an enzyme immunoassay, when a specific antibody produced by contacting a peptide essential to the formation of a specific antibody to a peptide antigen, a freeze-dried material of .beta.-D-galactosidase-enzyme conjugate or a peptide-enzyme conjugate prepared by coupling a labeling enzyme with a peptide of the general formula:H-R.sub.1 -Pro-Ser-Asp-Thr-Pro-Ile-Leu-Pro-Gln-OHwherein R.sub.1 is a peptide fragment consisting of 1 to 14 amino acid residues including Gly in the 14-position of the peptide Ala.sup.1 -Pro.sup.2 -Pro.sup.3 -Pro.sup.4 -Ser.sup.5 -Leu.sup.6 -Pro.sup.7 -Ser.sup.8 -Pro.sup.9 -Ser.sup.10 -Arg.sup.11 -Leu.sup.12 -Pro.sup.13 -Gly.sup.14 is used, a high reproducibility of the result of the enzyme immunoassay is obtained.

Abstract: There is described an assay for determining the relative concentrations of two antigenic solutes in a sample. The assay involves a first antibody to the first antigenic solute, a second antibody to the second antigenic solute and a ligand molecule having substituents to which the first and second antibodies may bind. The second antibody is immobilized upon a solid phase support and the other components are in solution. The components are mixed with a sample containing the antigenic solutes causing competition between the ligand molecule and each of the antigenic solutes for binding to the first and second antibodies. In the situation where there is a relatively higher concentration of the first antigenic solute to the second antigenic solute in the sample, a significant number of the ligand molecules become attached to the solid phase via the second antibody yet are capable of binding to a labelled antibody having the same specificity as the first antibody.

Abstract: Method and kit for pregnancy detection adapted for self-performance. Urine to be tested is contacted with a lectin substrate being lectin bound to a solid support and capable of binding HCG. After separation of the lectin substrate from the urine the substrate is contacted with a liquid color reagent comprising a colored carrier material and anti-HCG antibodies bound thereto. If after separation of the colored reagent from the substrate the latter remains colored, the subject is pregnant, while lack of color indicates non-pregnancy. A preferred color reagent comprises killed and stained Staphylococci bacteria. There is also provided a kit for self-performance of the method comprising at least one column packed with a lectin substrate and adapted for the controlled passage of liquid therethrough, and a color reagent.

Abstract: In a method for immunochemical assay of human chorionic gonadotropin involving use of an antibody immobilized on a carrier, an antigen and an antibody to which a labeling agent has been attached, when the antibody supported on the carrier and the antibody coupled to a labeling agent are different antibodies which are not overlapping in antigen-determining site and one of said different antibodies is specifically reactive to human chorionic gonadotropin, a high reproducibility of the result of the immunochemical assay is obtained.

Abstract: The present invention relates to a novel method for the quantitative determination of PEP (progestagen-associated endometrial protein) in a body fluid by radioimmunoassay. The technique is useful in monitoring the function of the human endometrium.

Abstract: This invention relates to a highly-sensitive mini-iodinated radioactive hormone tracer which has both initial low iodine-to-hormone molar ratios and low specific activity. This invention also relates to methods for preparing and using such tracers in a radio-immunoassay which takes much less time than used heretofore and to a method of preparing said tracers.

Abstract: A method is provided for determining the concentration of pregnanediol glucuronide (PG) in a woman's urine which is characterized by utilization of the reagent 20.alpha.-hydroxy-4-pregnen-3-one (20-.alpha. reagent) in a form in which it reacts with antibodies binding to PG. The method is adaptable to a visual color indication test which can be performed by the woman herself as well as by laboratory analysis. The method can be used to define the period in which conception can occur, to define a post-ovulation safe period in which conception is prevented, and as an early pregnancy indicator.

Abstract: Reagents and methods are described for an immunoassay test of increased sensitivity and decreased complexity employing an immunoglobulin specific for an antigen naturally or artificially placed upon the surface of an indicator particle coupled through the use of a hetero-bifunctional coupling reagent to a second antibody of differing specificity and specific for the antigen to be detected. In a preferred embodiment, a hetero-bifunctional coupling agent couples via a sulfhydryl group, a univalent immunoglobulin specific for the surface antigens on erythrocytes to a second multivalent immunoglobulin through an amide linkage with the latter immunoglobulin wherein said second immunoglobulin is specific for hepatitis-B surface antigen.

Abstract: Latex agglutination test methods and kits. Employing dyed latex polymer particles and adding a water-soluble, non-latex polymer particle absorbing dye contrasting in color to the dyed latex polymer particles produces a reaction mixture that changes color when agglutination occurs. Direct and indirect latex agglutination tests are contemplated which are useful in detecting disease states or physiological states such as pregnancy.

Abstract: The sensitivity of hemagglutination inhibition tests is improved by introducing a determined amount of lyophilized antigen or antibody into serological tubes in the absence of the indicator component. The test fluid to be analyzed is incubated solely in the presence of its binding partner in a liquid phase. After completion of the binding reaction (about 5 hours but extendable to 18 hours), the sensitized indicator solid phase, usually consisting of sensitized red blood cells, is added. By this process a 10 to 20 fold increase in sensitivity of the hemagglutination inhibition test is routinely achieved.

Abstract: A method is provided for determining the concentration of pregnanediol glucuronide (PG) in a woman's urine which is characterized by utilization of the reagent 20.alpha.-hydroxy-4-pregnen-3-one (20-.alpha. reagent) in a form in which it reacts with antibodies binding to PG. The method is adaptable to a visual color indication test which can be performed by the woman herself as well as by laboratory analysis. The method can be used to define the period in which conception can occur, to define a post-ovulation safe period in which conception is prevented, and as an early pregnancy indicator.