Enzyme-Linked Immunosorbent Assay (ELISA) is a widely used analytical technique in equine research for detecting and quantifying specific proteins, hormones, and antibodies in horse biological samples. This method relies on antigen-antibody interactions and employs enzyme-linked antibodies to produce a measurable signal, typically a color change, indicating the presence and concentration of the target molecule. ELISA is applicable in various areas of equine health, including the diagnosis of infectious diseases, monitoring of immune responses, and assessment of physiological conditions. It is valued for its specificity, sensitivity, and ability to process multiple samples simultaneously. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and advancements of ELISA in equine science.
Evans KS, Carpenter SL, Sevoian M.The enzyme-linked immunosorbent assay (ELISA) antigen-positive and agar-gel immunodiffusion test (AGID)-negative horses do not have infective equine infectious anemia (EIA) virus. The ELISA testing of horse leukocyte culture (HLC) supernatants did detect EIA virus in a HLC that was infected with the Wyoming strain of EIA virus and in HLC derived from horses in febrile, acute, or subacute stages of EIA infection. In supernatants of HLC derived from chronic and inapparent carrier horses, EIA virus was not detected with ELISA. Direct fluorescent antibody tests detected EIA virus in HLC infected w...
Suter M, Fey H.Horse IgE was isolated from a serum pool collected from foals naturally infected with endoparasites. The serum was precipitated with ammonium sulfate, delipidated with dextran sulfate and further purified by gel filtration, anionic exchange, immunosorption or preparative polyacrylamide gelelectrophoresis. By these methods IgE could be isolated at a purity of 81%. The sera from rabbits immunized with the purified horse serum fractions were tested using reversed passive cutaneous anaphylaxis and an enzyme linked immunosorbent assay (ELISA). By the ELISA method cross reaction of rabbit anti horse...
Suter M, Fey H.An enzyme-linked immuno sorbent assay (ELISA) for measuring horse IgE specific to ovalbumin, bencylpenicilloic acid and odinitrocarboxyphenol is described. We used a sandwich type of ELISA by which horse serum was incubated in antigen-coated tubes containing one additional polystyrene ball, followed by rabbit anti horse IgE serum. The tubes were then incubated with biotinylated goat anti rabbit globulin followed by avidin coupled to phosphatase. Endpoint titrations were compared. The ELISA is highly reproducible due to the pretreatment of the polystyrene with glutaraldehyde. The increased anti...
Braverman Y, Ungar-Waron H, Frith K, Adler H, Danieli Y, Baker KP, Quinn PJ.A survey of sweet itch in horses in Israel based on a questionnaire to owners reported that 158 of 723 horses (21.8 per cent) had sweet itch lesions. The results indicated that the likelihood of a horse acquiring sweet itch decreased with increasing altitude but no definite association with rainfall zones was evident. Variation in the density of the horse population, however, obscured these observations. In the population surveyed, stallions were more sensitive than mares and pale horses appeared to be less sensitive than dark ones, but the sample size of this latter group was much smaller. In...
Suzuki T, Ueda S, Samejima T.An enzyme-linked immunosorbent assay (ELISA) was elaborated for the detection of specific antibody to equine infectious anemia (EIA) antigen. Sera from horses experimentally infected with EIA virus were assayed by ELISA, complement fixation (CF) and immunodiffusion (ID) tests for antibody to EIA antigen. The ELISA technique was found to be much more sensitive than CF and ID tests. In addition, EIA specific antibody could be detected by ELISA at an earlier stage of infection than by CF or ID techniques. The applicability of the technique to diagnosis of EIA is discussed.
Kirchhoff H, Ammar AM, Heitmann J, Dubenkropp H, Schmidt R.Sera from horses with respiratory disease (RD) have been investigated using the complement fixation test, indirect hemagglutination test, enzyme immune assay, and the metabolic inhibition test, and sera from mares after abortion, using the complement fixation test, indirect hemagglutination test and enzyme immune assay, for antibodies against Mycoplasma equirhinis, M subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon and A. equifetale. Antibodies were found against all mycoplasma and acholeplasma species tested, more often against acholeplasmas. The antibod...
Bártek J, Viklický V, Franĕk F, Angelisová P, Dráber P, Jarosíková T, Nĕmec M, Verlová H.Five stable hybridoma lines were prepared using the myeloma cell line P3-X63-Ag.653 and spleen cells of mice hyperimmunized by pig transferrin. All hybridomas grew well in mouse peritoneal cavity and produced antibodies of the IgG1 subclass. Antibody preparations obtained from ascitic fluids tested for their capacity of antigen precipitation. No precipitation was obtained with single antibodies and with pairs of antibodies. Three out of 10 possible triads gave clear and sharp precipitation zones and rings in immunodiffusion tests performed in agar gel. All 5 antibodies were shown by quantitati...
Ueda Y, Sanai Y, Homma JY.Enzyme-linked immunosorbent assay (ELISA) was used for detection of immunoglobulin (Ig) M and IgG antibodies against a serologically common antigen (original endotoxin protein), protease, and elastase of Pseudomonas aeruginosa. The P aeruginosa antibody in horse sera was measured, using ELISA. Horseradish peroxidase-labeled rabbit anti-horse IgM and IgG antibodies were used for enzyme-labeled antibody conjugate. 5-Aminosalicylic acid and H2O2 were used for substrate. Sera collected from a vaccinated horse, a newborn foal, and 72 healthy racehorses were investigated for antibodies against P aer...
Sahu SP.Pony mares were vaccinated with killed contagious equine metritis (CEM) bacteria by IV, subcutaneous, and intrauterine (IU) routes (or a combination of these routes). The serum agglutinating antibody titer varied from 1:64 to 1:1,024 after vaccination. In pony mares challenge exposed with 96-hour-old culture of CEM bacteria given by IU route, there were clinical signs of CEM, but these signs were less severe in vaccinated mares than in nonvaccinated mares. The bacterium was isolated for the exudate and from uterine samples collected from the mares after challenge exposure. A low titer of IU an...
Potgieter LN, Rouse BT, Webb-Martin TA.A modification of the indirect enzyme-linked immunosorbent assay (ELISA) was developed which used staphylococcal protein A linked to horseradish peroxidase. Virus antibodies in equine, bovine, porcine, feline, canine, lagomorphic (rabbit), and human sera were detected, using the indirect ELISA in which the antiglobulin enzyme conjugate was replaced by protein A linked to horseradish peroxidase. Results of the ELISA were compared with the results of the serum-virus neutralization test. The application of the test in laboratories performing serologic assays with sera from diverse animal species ...
Ammar AM, Heitmann J, Kirchhoff H.After abortion sera were taken from 58 thoroughbred and other mares of the northwestern part of Germany and investigated by ELISA (enzyme linked immuno-sorbent assay) for antibodies against Mycoplasma equirhinis, M. subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon, and A. equifetale. Reactions at serum dilutions of 1:32 and higher were considered as positive. At serum dilution 1:32 no antibodies were found in 11 sera. The remaining sera showed antibodies against one or more of the mycoplasma antigens investigated. The number of multiple reactions decrease...
Kenney RM, Cummings MR, Teuscher C, Love CC.Testicular degeneration is a major cause of subfertility in stallions, although an aetiological diagnosis cannot be made in most cases. In the present study, autoimmune testicular degeneration was induced and evaluated in stallions by immunizing stallions with their own spermatozoa mixed with an adjuvant. The factors evaluated included changes in semen quality and testicular histology. A large decrease in sperm number and quality was observed in response to sperm autoantigens. An ELISA test specific for antisperm antibodies was developed which enabled antibody titres in serum, seminal plasma a...
Corstvet RE, Gaunt SD, Karns PA, Burgermeister D, McBride JW, Nicholson SM, Battistini RA.Four horses were inoculated with Ehrlichia risticii contained in either infected murine P388 D1 cells or heparinized blood from an infected horse. All 4 horses produced serum antibody, plasma antigen, and clinical signs of the disease. An enzyme-linked immunosorbent assay was used to detect antibody in the serum and was also used in conjunction with an anti-E. risticii monoclonal antibody to detect antigenemia. These laboratory and clinical findings were correlated to determine the efficiency of the antigen detection method for discerning E. risticii infection.
McDonald J, Gall R, Wiedenbach P, Bass VD, DeLeon B, Brockus C, Stobert D, Wie S, Prange CA, Ozog FJ.A one step enzyme-linked immunosorbent assay (ELISA) test for morphine was evaluated as part of a panel of pre- and post-race tests for narcotic analgesics in racing horses. This ELISA test is very sensitive to morphine with an I-50 for morphine of about 400 pg/ml. The test is also rapid, and ten samples, a normal pre-race complement, can be analyzed in about thirty minutes. The test can be read with an inexpensive spectrophotometer, or even by eye. The test readily detects the presence of morphine or its metabolites in equine blood for up to six hours after administration of sub-therapeutic d...
Smith ML, Chapman CB.An acepromazine (ACP) hapten was synthesised, coupled to bovine serum albumin and injected into a horse to produce antibodies to the drug. A competitive ELISA was developed whereby ACP attached to the solid phase via lysozyme competed with free ACP present in phosphate buffered saline, horse serum or horse urine for limiting amounts of antibody. The assay could detect the presence of ACP and, or, some of its metabolites in horse urine for at least 25 hours after intravenous injection of 0.1 mg kg-1 ACP maleate, but because of non-specific interference, horse serum could not be used. As little ...
Magnarelli LA, Ijdo JW, Van Andel AE, Wu C, Oliver JH, Fikrig E.To test serum samples of dogs and horses by use of class-specific recombinant-based ELISA for establishing a diagnosis of granulocytic ehrlichiosis attributable to infection with organisms from the Ehrlichia phagocytophila genogroup. Methods: Serum samples from 43 client-owned dogs and 131 horses (81 with signs of acute illness and 50 without signs of disease). Methods: Serum samples were analyzed, using ELISA with a recombinant 44-kd protein antigen for IgM and IgG antibodies to the human granulocytic ehrlichiosis (HGE) agent (NCH-1 strain). Western blot analyses, using infected human promyel...
Gaĭdamovich SIa, Pomelova VG, Lavrova NA, Mel'nikova EE, Sokolova MV, Kharitonenkov IG, Zlobin VN.Potentialities of differentiation between Venezuelan equine encephalomyelitis (VEE) complex viruses by time-resolved fluoroimmunoassay and enzyme immunoassay were studied. For this, 4 test systems were used based on different combinations of native and labeled polyclonal antibodies to VEE virus, strain Trinidad, and monoclonal (MCA) antibody MAK 14-7 to protein EL of this virus. The maximal sensitivity and specificity was achieved in the test system formed from native MCA MAK 14-7 for sensitization of the solid phase and labeled polyclonal immunoglobulins for demonstration of the test results....
Chen JW, Uboh CE, Robinson MA, Jiang Z, Soma LR.To evaluate plasma interleukin 6 (IL-6) concentration in Standardbred racehorses by means of a novel ELISA following validation of the assay for use with equine plasma samples. Methods: Plasma samples obtained from 25 Thoroughbreds for use in assay validation and from 319 Standardbred racehorses at rest 2 to 2.5 hours prior to warm-up and racing. Methods: A sandwich ELISA was developed with equine anti-IL-6 polyclonal antibody and the biotin-streptavidin chemical interaction to enhance sensitivity. The assay was validated for specificity, sensitivity, precision, and accuracy by use of both rec...
Vianna AM, Santos AC, Cunha RC, de Souza Stori de Lara AP, Weege GB, Leivas Leite FP.Equine theileriosis is a severe equine disease caused by the protozoan Theileria equi, which is prevalent in tropical and subtropical areas. In this study, a recombinant equi merozoite antigen-2 (rEMA-2) of T. equi was used as an immunogen. Two groups of 10 mice each were divided into control and vaccinated groups. Sixty mares seronegative for theileriosis were divided in two groups, one vaccinated and another group as a control animal. Mice and mares of the vaccinated groups were inoculated with 150 μL of the vaccine containing 50 μg of rEMA-2 and 2 mL of the vaccine containing 200 ...
Lygren T, Hansen S, Langberg H, Fjeldborg J, Jacobsen S, Nielsen MO, Schjerling P, Markussen B, Thomsen PD, Berg LC.Insulin-like growth factor 1 (IGF-1) has important roles in anabolic processes in the musculoskeletal system and has been reported to decrease with age in both people and horses. Objective: The objective of this study was to determine serum IGF-1 levels in the aging horse from early to late adulthood (age range 5-27 years). Methods: Healthy horses (n = 72) were used in a cross-sectional study, while 37 paired serum samples were available for a longitudinal study. Serum IGF-1 protein was determined using an ELISA kit validated for use in equine samples. Results: No association was found betw...
Velineni S, Timoney JF, Artiushin SC, Donahue JM, Steinman M.Foals of mares infected with Leptospira interrogans serovar Pomona type kennewicki (Lk) may be aborted/stillborn or delivered as healthy foals. Is fetal survival explained in part by the immune response of the fetus to Leptospira antigens? Objective: To describe an outbreak of Leptospira abortion in which infected mares delivered dead/sick or normal foals and determine specificities of antibody in a collection of 54 fetuses from similar outbreaks. Methods: Outbreak investigation in combination with a case-control study of a larger set of samples from aborted fetuses. Methods: Serology and poly...
Dyke TM, Sams RA.The objective of this study was to determine the urinary excretion of methylxanthines in horses following ingestion of chocolate over eight days. The study was performed in response to gas chromatography-mass spectrometry (GC-MS) confirmation of the presence of caffeine in a positive urine test in a racehorse. The trainer of the horse alleged that he often administered chocolate-coated peanuts as treats to his horses, and he believed that the ingestion of chocolate was responsible for the positive urine test. The urinary excretion of theobromine and caffeine after the ingestion of chocolate-co...
Benoit M, Lingen K, Taddei LM, Heffron BT, Hurt L, Lokanc JA, Lingner K, Cardenas E, Flores S, Mayer D, Pilipiak D, Folker-Calderon D, Negrusz A.Pyrilamine (mepyramine) is an H1-receptor antagonist used in human and veterinary medicine. It has the potential to produce central nervous system effects in horses and therefore may have some impact on an outcome of a horse race. A single oral dose of pyrilamine (300 mg/horse) was given to three animals. Serum samples were collected before drug administration and at 0.25, 0.5, 1, 2, 4, 6, 24, 48, 72, 96, 120, and 144 h, and 7, 8, 9, 10, 11, 12, and 13 days post-administration. Urine samples were collected at 0-1, 1-2, 2-4, 4-6, 24, 48, 72, 96, 120, and 144 h, and 7, 8, 9, 10, 11, 12, 13 days ...
Rabinovsky ED, Yang TJ.A tumor antigen (TA) associated with murine leukemia-lymphoma L5178Y cells has been identified by the enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF) techniques. The antigen was present in both non-solubilized and 0.5% NP-40 solubilized membrane extracts. Rabbit anti-L5178Y lymphoma serum (RALS), extensively absorbed with normal mouse tissues, identified TA in extracts of L5178Y lymphoma and L5178Y leukemia cells grown in horse serum (L5178Y/HS), but not in extracts of L5178Y cells grown in fetal calf serum (L5178Y/FCS). Similarly, absorbed rabbit anti-L5178Y/HS...
