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.
Abutarbush SM, Alqawasmeh DM, Mukbel RM, Al-Majali AM.The purposes of this study were to estimate the seroprevalence and distribution of horse piroplasmosis, to evaluate risk factors associated with the occurrence of the disease and to compare the different diagnostic methods used for this disease. A total of 253 clinically normal horses were sampled, and a collection form was completed for each horse from five of six different climatic zones of Jordan. The sixth zone was not sampled because it did not include horse population. Competitive enzyme-linked immunosorbent assay (cELISA) revealed 37 horses (14.6%) positive for Theileria equi, and none ...
Kumar S, Malik P, Verma SK, Pal V, Gautam V, Mukhopadhyay C, Rai GP.Glanders, caused by the Gram-negative, nonmotile bacterium Burkholderia mallei, is a contagious and highly fatal disease of equines. During the last decade, the number of glanders outbreaks has increased steadily. The disease also has high zoonotic significance and B. mallei is listed biological warfare agent. The complement fixation test (CFT) is a routinely used and internationally recognized test to screen equine sera for the glanders. However, discrepant results have been observed using the CFT. The low sensitivity and specificity of the CFT and enzyme-linked immunosorbent assay (ELISA) ha...
Pauvolid-Corrêa A, Morales MA, Levis S, Figueiredo LT, Couto-Lima D, Campos Z, Nogueira MF, da Silva EE, Nogueira RM, Schatzmayr HG.Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for ...
Read AJ, Finlaison DS, Gu X, Davis RJ, Arzey KE, Kirkland PD.During the equine influenza (EI) outbreak, two assays were used in parallel to diagnose the disease, to demonstrate freedom from infection in disease control zones and ultimately to demonstrate that EI virus had been eliminated from the Australian horse population. A longitudinal study of a population of naturally infected horses was established to determine the performance characteristics of these assays.
Kirkland PD, Delbridge G.An important consideration in the selection of a vaccine during the Australian equine influenza (EI) outbreak in 2007 was the ability to differentiate between infected and vaccinated animals (DIVA). A blocking enzyme-linked immunosorbent assay (bELISA) targeted for the nucleoprotein of influenza A viruses was developed to differentiate between naturally infected horses and horses vaccinated with the ProteqFlu® vaccine, which only induces a response to viral haemagglutinin. This bELISA assay met the DIVA requirements and was used extensively during the EI control and eradication programs and '...
Sergeant ES, Cowled BD, Bingham P.This observational study was undertaken in order to evaluate the diagnostic specificity of the blocking enzyme-linked immunosorbent assay (bELISA) for serum antibodies to influenza A virus nucleoprotein during the equine influenza (EI) outbreak response in New South Wales, Australia, in 2007. Using data collected during the outbreak response, bELISA testing data were collated for assumed uninfected horses from areas where EI infection was never recorded. Diagnostic specificity of the bELISA used during the EI response was high, but varied significantly between some regions, although the reason...
Richard EA, Pitel PH, Christmann U, Lekeux P, Fortier G, Pronost S.Surfactant protein D (SP-D), mainly synthesised by alveolar type II cells and nonciliated bronchiolar cells, is one important component of innate pulmonary immunity. In man, circulating concentrations of SP-D are routinely used as biomarkers for pulmonary injury. To date, serum SP-D levels have only been investigated in horses in an experimental model of bacterial airway infection. Objective: To compare serum SP-D concentrations at rest and after exercise in horses with and without inflammatory airway disease (IAD). Methods: Venous blood samples were collected from 42 Standardbred racehorses a...
Tinworth KD, Wynn PC, Boston RC, Harris PA, Sillence MN, Thevis M, Thomas A, Noble GK.Determining circulating equine insulin concentrations is becoming increasingly important in equine clinical practice and research. Most available assays are optimized for human medicine, but there is strong equine cross-reactivity because of the highly conserved nature of insulin. To identify an accurate and reliable assay for equine insulin, 6 commercial immunoassays were evaluated for precision, accuracy, and specificity. Only 1 assay initially reached the requisite standard: Mercodia Equine Insulin Enzyme-linked Immunosorbent assay (ELISA). Plasma matrix interferences were identified when t...
Lejeune JP, Sandersen C, Votion D, Caudron I, Vander Heyden L, Franck T, Ceusters J, Mouithys-Mickalad A, Niesten A, De La Rebière de Pouyade G....Intensive exercise induces a systemic inflammatory response characterised by an increase of blood neutrophil count and myeloperoxidase (MPO) release. Neutrophil elastase (NE) could also contribute to tissues lesions by its proteinase activities. Objective: To compare plasmatic NE concentrations before and after different forms of intensive exercise. Methods: EDTA blood samples were taken from 51 eventing horses (EvH) and 32 endurance horses (EndH) were sampled before the race (T0). Blood sampling was performed 2 h (T1) after completing either phase D of a one or 2 star eventing competition (n ...
Ferlazzo A, Medica P, Cravana C, Fazio E.Involvement of thyroid function on performance warrants further investigation as limited data exists on the effects of showjumping on the dynamics of total and free iodothyronines. Objective: To investigate the response of circulating total and free iodothyronines in horses to experimental showjumping sessions and compare with the effects normally induced by competition and determine if fence height has any effect. Methods: Using a randomised crossover study design 6 trained horses were studied during experimental showjumping sessions over 10 fences of different height: 1.00 m (Session 1), 1.1...
Holbrook TC, McFarlane D, Schott HC.The inflammatory and neuroendocrine response to endurance exercise and relationship of these parameters to performance is not well documented in horses. HYPOTHESES OR OBJECTIVES: Evidence of systemic inflammation is associated with poor performance in horses competing in endurance events. Methods: Blood was collected prior to and at the finish or elimination point from horses competing in both the 80 and 160 km American Endurance Ride National Championship competitions in 2006. Immunoreactive alpha-melanocyte stimulating hormone (α-MSH) and tumour necrosis factor-alpha (TNF-α) were quantifie...
Liu X, Liu Q, Feng X, Tang Q, Wang Z, Li S, Feng Z, Zhu J, Guan X.Due to the disadvantages of human and equine rabies immunoglobulin, it is necessary to develop a substitute for HRIG and ERIG, especially for those people living in the developing countries. Because of higher affinity and lower immunogenicity of rabbit's immunoglobulins, anti-rabies immunoglobulins specific to rabies virus were produced in rabbits as a bioreactor, and had been characterized by ELISA, affinity assay, immunofluorescence assay (IFA), immunocytochemistry, rapid fluorescent focus inhibition test (RFFIT). ELISA, affinity assay and IFA showed that rabbit RIG (RRIG) bound specifically...
Boly R, Dessy S, Kohnen S, Kini F, Lompo M, Mouithys-Mickalad A, Guissou IP, Dubois J, Deby-Dupont G, Serteyn D, Franck T.Agelanthus dodoneifolius DC Danser (Loranthaceae) is used for the treatment of various diseases including asthma. The aqueous and hydroalcoholic extracts have been reported to have anti-inflammatory, spasmolytic and bronchorelaxant activities. The present study investigates the effects of the aqueous decoction and the diethyl ether, ethyl acetate and butanolic fractions of Agelanthus dodoneifolius DC Danser (Loranthaceae) on reactive oxygen species (ROS) production and myeloperoxidase (MPO) release by phorbol 12-myristate 13-acetate (PMA)-stimulated equine neutrophils and on purified equine MP...
Kranenburg LC, van Ree HE, Calis AN, de Pater M, Buter GJ, van Maanen C, Sloet van Oldruitenborgh-Oosterbaarn MM.Equine proliferative enteropathy caused by Lawsonia intracellularis is an emerging disease of weanling foals and affects their growth and development. The prevalence of Lawsonia intracellularis in The Netherlands is not known. The aim of the study was to investigate the seroprevalence of Lawsonia intracellularis in horses in The Netherlands. Blood samples were taken from healthy foals before and after weaning and from healthy yearlings and mature horses on farms throughout The Netherlands. These samples were analysed for the presence of Lawsonia intracellularis-specific antibodies with a block...
Ji Y, Guo W, Zhao L, Li H, Lu G, Wang Z, Wang G, Liu C, Xiang W.An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for the detection of the equine influenza virus (EIV), employing monoclonal and polyclonal antibodies against the A/equine/Xingjiang/2007 (H3N8) nucleoprotein (NP). Immunoglobulin G antibodies were purified and used as capture or detector antibodies. The specificity of the optimized AC-ELISA was evaluated using EIV, equine herpesvirus 1 (EHV-1), equine herpesvirus 4 (EHV-4), equine arteritis virus (EAV) and Japanese encephalitis virus (JEV), resulting in only EIV specimens yielding a strong signal. A minimal concentr...
Crafford JE, Guthrie AJ, Van Vuuren M, Mertens PP, Burroughs JN, Howell PG, Batten CA, Hamblin C.A polyclonal antibody-based, group-specific, competitive ELISA (C-ELISA) for the detection of antibodies to equine encephalosis virus (EEV) was developed. The assay measures the competition between a specific guinea pig antiserum and a test serum, for a pre-titrated EEV antigen. The C-ELISA detected antibodies to the seven known EEV serotypes. Reference antisera raised against other arboviruses did not cross react with EEV antigen. Negative sera from horses in the United Kingdom were used to establish the baseline for a negative population. Negative and positive populations of South African ho...
Baldani CD, Hilario E, Nakaghi AC, Bertolini MC, Machado RZ.The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in Escherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the E...
West Nile virus (WNV), the most widely distributed flavivirus worldwide, has lately reemerged in Europe, causing worrisome outbreaks in humans and horses. Serological analysis by enzyme-linked immunoassay and plaque reduction neutralization test showed for the first time in Serbia that 12% of 349 horses presented specific neutralizing WNV antibodies, which in one case also cross-neutralized Usutu virus (USUV). This is the first time that anti-USUV high neutralizing antibody titers are reported in horses. All these data indicate that WNV and USUV are circulating in the region and advise on the ...
Hraha TH, Doremus KM, McIlwraith CW, Frisbie DD.Osteoarthritis (OA) is one of the most prevalent and debilitating conditions affecting the horse. Autologous conditioned serum (ACS), commercially available as IRAP and IRAP II, is a recently developed treatment for OA in which plasma is prepared from venous blood by incubation with glass beads for 24 h. This product has been shown to increase anti-inflammatory cytokines and growth factors in human blood. However, data for equine ACS preparations are lacking. Objective: To characterise the protein profiles produced by commercially available ACS systems in equine blood. Methods: Blood was dra...
Dangoudoubiyam S, Oliveira JB, Víquez C, Gómez-García A, González O, Romero JJ, Kwok OC, Dubey JP, Howe DK.Serum samples from 315 horses from Costa Rica, Central America, were examined for the presence of antibodies against Sarcocystis neurona, Neospora spp., and Toxoplasma gondii by using the surface antigen (SAG) SnSAG2 enzyme-linked immunosorbent assay (ELISA), the NhSAG1 ELISA, and the modified agglutination test, respectively. Anti- S. neurona antibodies were found in 42.2% of the horses by using the SnSAG2 ELISA. Anti- Neospora spp. antibodies were found in only 3.5% of the horses by using the NhSAG1 ELISA, and only 1 of these horses was confirmed seropositive by Western blot. Antibodies to T...
Textor JA, Norris JW, Tablin F.To test the hypotheses that preparation method, exposure to shear force, and exposure to collagen affect the release of growth factors from equine platelet-rich plasma (PRP). Methods: PRP obtained from 6 horses. Methods: PRP was prepared via 2 preparation methods (tube and automated) and subjected to 6 treatment conditions (resting, detergent, exposure to shear via 21- and 25-gauge needles, and exposure to collagen [10 and 20 μg/mL]). Concentrations of platelet-derived growth factor, isoform BB (PDGF-BB); transforming growth factor β, isoform 1 (TGFβ₁); and insulin-like growth factor, iso...
Kitai Y, Shirafuji H, Kanehira K, Kamio T, Kondo T, Konishi E.West Nile virus (WNV) and Japanese encephalitis (JE) virus are distributed separately in the world with some exceptions. There is a concern that WNV may invade into Asia where JE virus exists. On and after such invasion, any differential diagnosis could be complicated by serological crossreactivities. We previously demonstrated experimentally using horses infected with WNV that preimmunization with inactivated JE vaccine considerably affected the ability of neutralization tests and immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (ELISA) to diagnose WNV infection. Here...
Sonmez K, Gurel A, Takai S.The aim of the present study was to develop an immunocytochemical procedure for the early detection and demonstration of Rhodococcus equi in smears of tracheal aspirates taken from live foals in field conditions. Tracheal wash samples were collected from thoroughbred foals, aged 1-5 months and located in studs around Bursa and Istanbul, Turkey. Some foals were suspected of having R. equi infection on the basis of clinical examination (n=56) and others were unaffected control animals (n=54). Serum samples were also collected from each foal for testing for the presence of R. equi-specific antibo...
Mujica FF, Perrone T, Forlano M, Coronado A, Meléndez RD, Barrios N, Alvarez R, Granda F.The main objective of this study was to demonstrate the occurrence of equine piroplasmosis (EP) in horses of Lara State, Venezuela, and to correlate it with the factors host's sex and age in order to know the epidemiology of this disease at the Venezuelan Centroccidental Region. Antibody levels to Babesia caballi and Theileria equi were assessed in 360 equine serum samples, collected from 9 municipalities of Lara State, using an ELISA technique with recombinant antigens and monoclonal antibodies (Mabs). Antibodies to B. caballi were found in 254 horses (70.6%), whereas 181 animals (50.3%) were...
Chang Y, Maylin GM, Matsumoto G, Neades SM, Catlin DH.Methods have been developed to screen for and confirm darbepoetin alfa, recombinant human EPO, and methoxy polyethylene glycol-epoetin β (PEG-epoetin β) in horse plasma. All three methods screen samples with an enzyme-linked immunosorbent assay (ELISA) and confirm by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This report focuses on PEG-epoetin β. The ELISA assay was able to detect PEG-epoetin β at 0.02 ng/mL in 50 µL of horse plasma. Many samples had high background levels of immunoreactivity; however, introducing polyethylene glycol 6000 (PEG 6000) into the samples before...
Ringger NC, Giguère S, Morresey PR, Yang C, Shaw G.Neonatal hypoxic-ischemic encephalopathy (NHIE) is a disease affecting newborn foals for which there is no antemortem diagnostic test. Objective: Ubiquitin C-terminal hydrolase 1 (UCHL1) and the phosphorylated axonal forms of neurofilament H (pNF-H) are markers of brain injury in foals with NHIE. Methods: Thirty-three foals with a clinical diagnosis consistent with NHIE and 17 healthy foals. Methods: Retrospective study. Concentrations of UCHL1 and pNF-H in plasma were measured by ELISA. The performance of the assays for the diagnosis of NHIE was assessed by receiver operating characteristic c...
Smith MR, Wright IM, Minshall GJ, Dudhia J, Verheyen K, Heinegård D, Smith RK.To evaluate digital flexor tendon sheath (DFTS) synovial fluid cartilage oligomeric matrix protein (COMP) concentrations as a molecular marker for intrathecal pathology. Methods: Case control study. Methods: Horses (n=46) with DFTS tenosynovitis; 23 fresh cadaver horses. Methods: DFTS synovial fluid samples were collected from clinical cases with noninfected DFTS tenosynovitis and from control DFTS. Clinical and surgical findings were recorded, and dissection of control limbs was performed to confirm the DFTS to be grossly normal. Synovial fluid COMP was quantified using a homologous competiti...
Yeargan MR, Howe DK.Equine protozoal myeloencephalitis (EPM) is a common neurologic disease of horses that is caused by the apicomplexan pathogen Sarcocystis neurona. To help improve serologic diagnosis of S. neurona infection, we have modified existing enzyme-linked immunosorbent assays (ELISAs) based on the immunogenic parasite surface antigens SnSAG2, SnSAG3, and SnSAG4 to make the assays polyvalent, thereby circumventing difficulties associated with parasite antigenic variants and diversity in equine immune responses. Two approaches were utilized to achieve polyvalence: (1) mixtures of the individual recombin...
Dart AJ, Dart CM, Dudhia J, Perkins N, Canfield P, Smith RK.To evaluate whether wound type or site influence the production of cartilage oligomeric matrix protein (COMP) and transforming growth factor β1 (TGF-β1), and determine if there is a correlation between TGF-β1and COMP during healing. Methods: Experimental. Methods: Standardbred horses (n=6), 4-8 years old. Methods: Six, standardized, full-thickness skin wounds (open, straight, and elliptical) were surgically created on the neck (n=3) and metacarpus (3) on each horse. Wounds were randomly allocated to site and side. Tissue samples were collected before creating wounds and on days 7, 14, and 4...
Salvagni CA, Dagnone AS, Gomes TS, Mota JS, Andrade GM, Baldani CD, Machado RZ.Ehrlichiosis is a zoonotic disease caused by gram-negative and intracellular obligatory bacterial organisms. Equine Granulocytic Anaplasmosis - EGA (formerly Equine Granulocytic Ehrlichiosis, EGE) is a seasonal disease, normally self-limited in horses. There are few reports in Brazil about this ehrlichial agent, as well as its natural vectors. Nowadays, veterinarians are considering the suspicion of EGA in horses with suggestive symptoms of ehrlichiosis and which do not respond to piroplasmosis treatment. The aim of the present study was to identify horses exposed to the agent A. phagocytophil...
Minervini F, Lacalandra GM, Filannino A, Nicassio M, Visconti A, Dell'Aquila ME.The purpose of this study was to assess the natural exposure of male horses (Equus caballus) to the mycotoxin zearalenone (ZEA) by using the ELISA test and to evaluate the effects of in vitro exposure of sperm cells to mycotoxin-containing urine extracts on sperm chromatin structure stability. Because of their occurrence in urine samples, ZEA and its derivatives were tested by sperm chromatin structure assay (SCSA) at natural levels detected by ELISA. Thirty-eight urine extracts of Italian (n = 11) and northeastern European (n = 27) horses were tested on frozen-thawed spermatozoa to evaluate t...
Ramos CP, Lopes EO, Oliveira Júnior CA, Diniz AN, Lobato FCF, Silva ROS.Considering the lack of studies evaluating the performance of commercially available methods for diagnosis of Clostridioides (Clostridium) difficile infection (CDI) in animals, the present study aimed to assess an immunochromatographic test for detection of glutamate dehydrogenase (GDH) and A/B toxins of C. difficile, also evaluated by an ELISA kit, in foals and neonatal piglets. Intestinal contents of 47 piglets and feces of 35 foals were tested to GDH antigen and A/B toxins in a lateral flow method (Ecodiagnostica, Brazil). Also, these samples were submitted to A/B toxin detection by an ELIS...
Laegreid WW, Skowronek A, Stone-Marschat M, Burrage T.There are three clinicopathologic syndromes associated with African horsesickness (AHS) virus infection in horses. These different forms of AHS (pulmonary, cardiac, and fever forms) vary in the organs affected, the severity of lesions, time of onset of clinical signs and mortality rates. We have studied the effects of infection with three cell culture passaged variants of AHS virus in naive North American horses. One of these viruses, AHS/4SP, consistently caused the pulmonary form of AHS with rapid onset of severe pulmonary edema and 100% mortality. A second variant, AHS/9PI, resulted in sign...
Berti A, Degl'Innocenti D, Stefani M, Liguri G, Ramponi G.A non competitive enzyme-linked immunosorbent assay (ELISA) specific for horse muscle acylphosphatase (E.C. 3.6.1.7.) has been developed. The purified anti-acylphosphatase antibodies were immobilized by passive absorption to a solid-phase support and incubated with known and unknown amounts of antigen. The antibody-acylphosphatase complex was quantified using the same antibody conjugated to horseradish peroxidase. The assay yields positive reactions with as little as 0.05 ng of antigen, with intra- and interassay coefficients of variation of 5% and 7%, respectively. On the basis of this assay ...
Mizushima D, Amgalanbaatar T, Davaasuren B, Molefe NI, Battur B, Battsetseg B, Inoue N, Yokoyama N, Suganuma K.Our previous studies report epidemics of non-tsetse-transmitted equine trypanosomosis in Mongolia. However, the current status of non-tsetse-transmitted equine trypanosomosis endemicity remains to be clarified in some parts of Mongolia. We previously reported the potential application of rTeGM6-4r-based diagnostic tools, an rTeGM6-4r-based immunochromatographic test (ICT) and an enzyme-linked immunosorbent assay (ELISA), in the serological surveillance of equine trypanosomosis in Mongolia. In the present study, the utility of the rTeGM6-4r-based ICT was validated. The rTeGM6-4r-based ICT accur...
Sofronic-Milosavljevic Lj, Ilic N, Djordjevic M, Savic M, Gruden-Movsesijan A, Cuperlovic K, Murrell KD.In the Balkan countries, where trichinellosis is a re-emerging zoonosis, it is of great importance to determine Trichinella infection prevalence among the major hosts, including horses. One method for monitoring prevalence is serological surveillance; however, the validity of serological methods in horses is not well understood. The dynamics of anti-Trichinella IgG production and circulating excretory/secretory (ES) antigens were investigated in three horses experimentally-infected with Trichinella spiralis. Horses were slaughtered at 32 week post infection (p.i.). Low worm burdens were found ...
Ellis GR, Daniels E.Direct electron microscopy (EM) and enzyme-immunoassay (rotazyme) results for the detection of rotaviruses in 346 enteric specimens from calves, lambs, piglets and foals were compared. The rotazyme test was at least 3 times more sensitive than direct EM in diagnosing infection. Rotavirus antigen was demonstrated by rotazyme in 22% of 280 scour samples and in 27% of 66 samples from non-scouring animals. There was an association between diarrhoea and higher amounts of rotavirus antigen. This prevalence of rotaviruses detected in animals with diarrhoea highlights the significant involvement of ot...
Lavoie-Lamoureux A, Maghni K, Lavoie JP.The systemic component of chronic inflammatory diseases may lead to clinical complications. High levels of TNFα, a pro-inflammatory cytokine, are found in human patients with COPD and asthma. Horses are also susceptible to an array of chronic inflammatory disorders possibly associated with systemic inflammation, including respiratory diseases. Currently, there is no commercially available ELISA validated to assess TNFα in equine serum samples. Moreover, the reported normal mean concentration of serum TNFα in horses vary greatly. Hence, we sought to optimize and validate a procedure to quant...
Onyiche TE, Sivakumar T, Tuvshintulga B, Nugraha AB, Ahedor B, Mofokeng L, Luka J, Mohammed A, Mbaya AW, Biu AA, Yokoyama N, Thekisoe O.Equine piroplasmosis is caused by apicomplexan parasites, namely, and , which are transmitted to equids principally through ticks. To ascertain the exposure of equines to agents of equine piroplasms, we tested serum samples collected from horses (n = 272) and donkeys (n = 170) in North-Western Nigeria for the presence of antibodies against and using IFAT and ELISA. The seroprevalence of in the horses determined using IFAT and ELISA was 48.89% and 45.96%, respectively, while for , it was 6.3% and 0.4%, respectively. For , the seroprevalence based on IFAT and ELISA results in donkeys was...
Durán-Ferrer M, Villalba R, Fernández-Pacheco P, Tena-Tomás C, Jiménez-Clavero MÁ, Bouzada JA, Ruano MJ, Fernández-Pinero J, Arias M....This study described the clinical, virological, and serological responses of immunologically naïve and vaccinated horses to African horse sickness virus (AHSV) serotype 9. Naïve horses developed a clinical picture resembling the cardiac form of African horse sickness. This was characterized by inappetence, reduced activity, and hyperthermia leading to lethargy and immobility-recumbency by days 9-10 post-infection, an end-point criteria for euthanasia. After challenge, unvaccinated horses were viremic from days 3 or 4 post-infection till euthanasia, as detected by serogroup-specific (GS) real...
Alzuheir I, Fayyad A, Jalboush N, Abdallah R, Abutarbush S, Gharaibeh M, Bdarneh M, Khraim N, Helal MA, Helal BA.West Nile fever (WNF) is a neurotropic, mosquito-borne disease affecting humans and domesticated animals, caused by a member of the genus . Over the last decades, this virus has been responsible for several cases of illness in humans and animals. The current epidemiological status of WNF in horses is insufficient, and in veterinarians, as an occupational hazard is unknown. This study aimed to investigate and determine the seroprevalence and risk factors for WNF in veterinarians and horses in Palestine. Methods: In this study, serum samples from 100 veterinarians and 87 horses were collected be...
Alvarez I, Gutierrez G, Ostlund E, Barrandeguy M, Trono K.We analyzed the performance of a single-band Western blot (WB) test using recombinant p26 (rp26) capsid protein of equine infectious anemia virus. According to the results obtained, the rp26 WB test is a reliable confirmatory diagnostic tool to be used as a complementary test after an enzyme-linked immunosorbent assay or agar gel immunodiffusion test yielding doubtful results.
Ji Y, Guo W, Zhao L, Li H, Lu G, Wang Z, Wang G, Liu C, Xiang W.An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for the detection of the equine influenza virus (EIV), employing monoclonal and polyclonal antibodies against the A/equine/Xingjiang/2007 (H3N8) nucleoprotein (NP). Immunoglobulin G antibodies were purified and used as capture or detector antibodies. The specificity of the optimized AC-ELISA was evaluated using EIV, equine herpesvirus 1 (EHV-1), equine herpesvirus 4 (EHV-4), equine arteritis virus (EAV) and Japanese encephalitis virus (JEV), resulting in only EIV specimens yielding a strong signal. A minimal concentr...
Piyasena TBH, Setoh YX, Hobson-Peters J, Prow NA, Bielefeldt-Ohmann H, Khromykh AA, Perera D, Cardosa MJ, Kirkland PD, Hall RA.In Australia, infection of horses with the West Nile virus (WNV) or Murray Valley encephalitis virus (MVEV) occasionally results in severe neurological disease that cannot be clinically differentiated. Confirmatory serological tests to detect antibody specific for MVEV or WNV in horses are often hampered by cross-reactive antibodies induced to conserved epitopes on the envelope (E) protein. This study utilized bacterially expressed recombinant antigens derived from domain III of the E protein (rE-DIII) of MVEV and WNV, respectively, to determine whether these subunit antigens provided specific...
Mankoc S, Hostnik P, Grom J, Toplak I, Klobucar I, Kosec M, Barlic-Maganja D.In the present study, a new one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) strategy with minor-groove-binder (MGB) technology for the detection of EAV from 40 semen samples of Slovenian carrier stallions was tested. A novel MGB probe (EAVMGBpr) and a reverse primer (EAV-R) based on the multiple sequence alignment of 49 different EAV strain sequences of the highly conserved ORF7 (nucleocapsid gene) were designed. The performance of the assay was compared with different molecular detection methods. Three different primer pairs targeting the ORF1b and ORF7 were used, ...
Alvarez I, Cipolini F, Wigdorovitz A, Trono K, Barrandeguy ME.The most used and reliable indicator of Equine infectious anemia virus (EIAV) infection is the detection of its specific antibodies in horse serum. In the present study, the performance of two commercial ELISA tests for the detection of EIAV antibodies as well as the potential advantages of their use as an EIAV infection screening tool were evaluated in 302 horse serum samples. Both ELISA assays showed 100% diagnostic sensitivity, and 92.3-94.3% diagnostic specificity. Discordant results were analyzed by immunoblot. The results showed that both ELISA tests are very efficient at detecting EIAV ...
Torfs SC, Maes AA, Delesalle CJ, Deprez P, Croubels SM.Serotonin is regularly measured in equine platelet-poor plasma in research settings. However, reported reference values vary between studies, partially because plasma serotonin concentrations are very low and a reliable and affordable detection method is lacking. A simple, rapid, and sensitive method for serotonin determination in equine platelet-poor plasma using liquid chromatography--tandem mass spectrometry (LC-MS/MS) was developed and validated. Results of a commercially available enzyme-linked immunosorbent assay (ELISA) were compared to the LC-MS/MS results, in order to validate a test ...
Gough MR, Mayhew G, Munroe GA.This paper tests the hypothesis that the local analgesic agent mepivacaine diffuses between adjacent equine synovial structures in the forelimb and with greater frequency than latex, gelatine dye or contrast media. We report the incidence of diffusion of mepivacaine between the distal interphalangeal joint (DIPJ) and navicular bursa (NB) of the forelimbs and between the intercarpal (IC) and radiocarpal (RC) joints of 31 fresh equine cadavers. The DIPJ of one forelimb and the NB of the contra lateral forelimb and the RC joint of one forelimb and the IC joint of the contra lateral forelimb were ...
Read AJ, Arzey KE, Finlaison DS, Gu X, Davis RJ, Ritchie L, Kirkland PD.An outbreak of equine influenza (EI) occurred in Australia in 2007. During the laboratory support for this outbreak, real-time reverse transcriptase polymerase chain reaction (qRT-PCR) assays and a blocking enzyme linked immunosorbent assay (bELISA) were used as testing methods to detect infection with the virus. The qRT-PCR and bELISA tests had not been used for EI diagnosis before, so it was not known how soon after infection these tests would yield positive results, or for how long these results would remain positive. To answer these questions, nasal swabs and blood samples were collected d...
Porter MB, Long M, Gosche DG, Schott HM, Hines MT, Rossano M, Sellon DC.The West Nile (WN) virus, present in the United States since 1999, is a cause of encephalomyelitis in birds, alligators, humans, and horses. No data exist regarding detection of anti-WN virus immunoglobins in equine cerebrospinal fluid (CSF). The aims of this study were to evaluate the blood-brain barrier (BBB) in WN virus-infected (WNE) horses, to compare diagnostic testing in serum and CSF, and to describe the immunoglobulin M (IgM) response in serum and CSF of vaccinated horses. CSF was collected from the lumbosacral (LS) space (n = 13) or the allanto-occipital (AO) space (n = 14) of WNE ho...
Alemu T, Luckins AG, Phipps LP, Reid SW, Holmes PH.A field study involving 309 horses was undertaken in the provinces of Arsi and Bale in the Ethiopian highlands to investigate the prevalence of Trypanosoma equiperdum infections using enzyme linked immunosorbent assays (ELISAs) for the detection of both trypanosomal antigen and antibody. Adult horses of both sexes were examined for clinical signs of T. equiperdum infection and serum samples were collected for the assays. One hundred and one horses showed the presence of trypanosomal antibodies in their serum and 70 animals showed typical clinical signs of dourine. Nineteen horses showed the pr...
Swadzba ME, Hirmer S, Amann B, Hauck SM, Deeg CA.Although the presence of IgG autoantibodies in the vitreous of spontaneous cases of equine recurrent uveitis (ERU) has been demonstrated, the potential role of IgM reactivities during ERU pathogenesis remains unexplored. The purpose of this study was to examine the presence of IgM autoantibodies in vitreous specimens of ERU-affected horses and to test their binding specificity to intraocularly expressed proteins. Methods: To test IgM autoantibody responses to retinal tissue, vitreous samples of eye-healthy controls and ERU patients were analyzed via two-dimensional Western blot analysis with e...
Cho HJ, Entz SC, Deregt D, Jordan LT, Timoney PJ, McCollum WH.A potent ELISA antigen was prepared from equine arteritis virus (EAV) by differential centrifugation of EAV-infected cell culture fluid, followed by solubilization of the preparation by Triton X-100 treatment. Using this antigen and a mouse monoclonal antibody against the G(L) protein of EAV, a reliable blocking ELISA (bELISA) was developed for the detection of EAV antibodies in equine sera. The bELISA was evaluated using a total of 837 test serum samples. The relative sensitivity (n = 320) of the bELISA compared to the serum neutralization (SN) test was 99.4%. The bELISA appears to be a highl...
Rhalem A, Sahibi H, Lasri S, Johnson WC, Kappmeyer LS, Hamidouch A, Knowles DP, Goff WL.A highly specific and sensitive competitive enzyme-linked immunosorbent assay for detection of specific antibody to Babesia equi in serum from equids was validated for use in Morocco. The assay is based on the specific inhibition of binding of a monoclonal antibody to a conserved epitope within a recombinant parasite peptide by serum from infected animals. The assay was compared to an established indirect immunofluorescence assay, with a concordance of 91%. The assay was used to determine seroprevalence for B. equi infections in donkeys and horses throughout Morocco. A total of 578 sera (163 h...
Schvartz G, Tirosh-Levy S, Barnum S, David D, Sol A, Pusterla N, Steinman A.Equine coronavirus (ECoV) infection is the cause of an emerging enteric disease of adult horses. Outbreaks have been reported in the USA, EU and Japan, as well as sporadic cases in the UK and Saudi Arabia. Infection of ECoV in horses in Israel has never been reported, and the risk of exposure is unknown. Importation and exportation of horses from and into Israel may have increased the exposure of horses in Israel to ECoV. While the disease is mostly self-limiting, with or without supportive treatment, severe complications may occur in some animals, and healthy carriers may pose a risk of infec...
Halassy B, Kurtović T, Lang Balija M, Brgles M, Tunjić M, Sviben D.The hyperimmune horse plasma (HHP), prepared through active immunisation of horses with an antigen of interest, is the most common starting material for antitoxin (animal antibody-based therapeutics) production. Precise IgG quantification in plasma is a prerequisite for accurate estimation of the purification process efficiency. Although immunoglobulins from HHP have been purified for over a century, there is still no in vitro method for precise and accurate determination of IgG content in HHP. For this reason, the purification process efficiency has been assessed by antibody activity measurem...
Waggett BE, McGorum BC, Wernery U, Shaw DJ, Pirie RS.While previous studies have demonstrated an association between equine grass sickness (EGS) and the presence of Clostridium botulinum within ileal contents and faeces, no such associations with other intestinal-derived anaerobic bacteria have been extensively investigated. Objective: The prevalence of C. perfringens in the ileal contents and faeces of EGS horses is greater than control horses; the detection of C. perfringens in faeces by ELISA could be diagnostically beneficial in a clinical setting. Methods: The prevalence of C. perfringens in faeces from EGS horses and healthy grazing contro...
Higuchi T, Hashikura S, Gojo C, Inui T, Satoh S, Yoshida M, Ishiyama T, Yamada H, Takai S.An enzyme-linked immunosorbent assay (ELISA) for detection of serum IgG antibodies against Tween 20-extracted antigen of strain ATCC 6939 was applied in Hidaka, Japan to a total of 752 sick foals showing a variety of signs of infectious disease. An optical density (OD) value of more than 0.3 was tentatively fixed to be positive on the basis of readings made of healthy horse sera in previous studies. During a 2 year study, 138 of the 752 sick foals showed an OD value of 0.3 or higher and were designated as 'suspected of R. equi infection'. Age distribution during the initial medical examination...
Kumar S, Kumar R, Gupta AK, Yadav SC, Goyal SK, Khurana SK, Singh RK.Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which ...
The National Reference Center for equine infectious anemia (EIA) validated a commercial ELISA (Eradikit EIAV Indirect ELISA, In3diagnostic, Turin, Italy) employing a chimeric recombinant gag and env peptide for the detection of EIA virus antibodies, following the guidelines of the World Organization for Animal Health. The validation parameters evaluated were: analytical sensitivity (Se) and specificity (Sp); diagnostic Se and Sp; precision, based on repeatability and reproducibility through the estimation of the standard deviation (SD) and the coefficient of variation (CV); accuracy, estimated...
