Topic:Serodiagnosis
Serodiagnosis in horses involves the detection and measurement of antibodies or antigens in the blood to diagnose infectious diseases and monitor immune responses. This diagnostic approach is based on serological tests that identify the presence of specific immune markers, providing insight into the horse's exposure to pathogens or vaccination status. Common serological tests used in equine medicine include enzyme-linked immunosorbent assays (ELISA), complement fixation tests, and virus neutralization tests. These tests are valuable for diagnosing conditions such as equine infectious anemia, strangles, and equine herpesvirus. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and advancements in serodiagnostic techniques for equine health assessment.
A field study to estimate the prevalence of Trypanosoma equiperdum in Mongolian horses. From May to July 2000, a cross-sectional study was conducted to estimate the prevalence of Trypanosoma equiperdum in the horse population of the central province (Tuv aimag) of Mongolia. On average, four herds were selected from each of the 29 aimag subdivisions (119 herds). From each herd, 10 horses were sampled in proportion to sex and age categories in the respective herds (1190 horses). Sera from 1122 horses were analysed for T. equiperdum antibodies using two serological assays, the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA). The crude estimate of the...
A recombinant envelope protein-based enzyme-linked immunosorbent assay for West Nile virus serodiagnosis. Recombinant West Nile virus envelope (E) protein was examined in enzyme-linked immunosorbent assay (ELISA) to detect antibodies elicited during West Nile virus infection. Horses (nine of 10) and humans (six of six) with confirmed West Nile virus infection had IgG and/or IgM antibodies to the E protein. Antibodies to the recombinant West Nile virus membrane and nonstructural 1 proteins were not detected in any of these sera. An E protein-based ELISA may aid in the serological diagnosis of West Nile virus infection.
Performance of five serological assays for diagnosis of Rhodococcus equi pneumonia in foals. The performance of four enzyme-linked immunosorbent assays (ELISAs) (ELISA-6939, ELISA-33701, ELISA-VapA, and ELISA-California) and an agar gel immunodiffusion test for diagnosis of Rhodococcus equi pneumonia in foals was evaluated. Antibody concentrations of foals with culture-confirmed R. equi pneumonia (n = 41) were compared to those of age-matched pasturemates that remained clinically healthy during the entire breeding season (n = 24). For each serological assay evaluated, selection of a low cutoff resulted in high sensitivity but low specificity. Increasing the cutoff value resulted in be...
High-level expression and purification of a truncated merozoite antigen-2 of Babesia equi in Escherichia coli and its potential for immunodiagnosis. The gene encoding a truncated merozoite antigen-2 (EMA-2t) of Babesia equi was cloned and highly expressed in Escherichia coli as a glutathione S-transferase fusion protein (G-rEMA-2t). Both G-rEMA-2t and rEMA-2t (after the removal of glutathione S-transferase) had good antigenicity. Either Western blot analysis with rEMA-2t or enzyme-linked immunosorbent assay (ELISA) with G-rEMA-2t clearly discriminated the sera of horses experimentally infected with B. equi from sera of horses infected with Babesia caballi and healthy horses, although rEMA-2t was not suitable for ELISA, probably owing to it...
A serological survey of Rhodococcus equi infection in foals in central Italy: comparison of two antigens using an ELISA test. A serological survey of Rhodococcus equi infection was carried out on 602 blood samples collected from foals in central Italy. The assay was performed with an ELISA test using two different antigens prepared with reference strains of R. equi, ATCC 33071 and ATCC 6939. A positive reaction was obtained on 81 serum samples (13.45%) (OD > or = 0.3) using antigen ATCC 33071, and on 73 serum samples (12.12%) using antigen ATCC 6939. Although the frequency of the disease was not high, the serological positivity was about 13%. There was no statistically significant difference between males and fema...
Standardisation and comparison of serial dilution and single dilution enzyme linked immunosorbent assay (ELISA) using different antigenic preparations of the Babesia (Theileria) equi parasite. Serial dilution and single dilution enzyme linked immunosorbent assays (ELISA) were standardised and their sensitivity and specificity were compared for serodiagnosis of Babesia equi infection. The antibody titres of 24 donkey sera of known identity were determined separately by serial dilution ELISA using three different B. equi antigens namely whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS). The ratios of the optical density (OD) of known positive and known negative sera at different serum dilutions were calculated and termed as the positive/negative (P/N) ratio. Th...
Comparison of a serum indirect fluorescent antibody test with two Western blot tests for the diagnosis of equine protozoal myeloencephalitis. A serum indirect fluorescent antibody test (IFAT) was compared with a Western blot (WB) and a modified Western blot (mWB) for diagnosis of equine protozoal myeloencephalitis (EPM). Using receiver-operating characteristic (ROC) analysis, the area under the curve of the IFAT was greater than the areaunder the curves of the WB and the mWB (P = 0.025 and P = 0.044, respectively). There was no statistically significant difference between the areas under the curves of the WBs (P > 0.05). On the basis of an arbitrarily chosen cut-off titer for a positive test result of 1:80 for the IFAT and interpret...
Identification of a specific antigenic region of the P82 protein of Babesia equi and its potential use in serodiagnosis. The efficacy of the Be82 gene product fused with glutathione S-transferase (GST/Be82) in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Babesia equi infection was reported previously (H. Hirata et al., J. Clin. Microbiol. 40:1470-1474, 2002). However, the ELISA with the GST/Be82 antigen cross-reacted with Babesia caballi-infected horse sera, despite the high rate of detection of B. equi. These results suggested that GST/Be82 has an antigen in common with B. caballi or antigenicity similar to that of B. caballi. In the present study, we constructed a series of five clones wit...
Experimental induction of equine protozoan myeloencephalitis (EPM) in the horse: effect of Sarcocystis neurona sporocyst inoculation dose on the development of clinical neurologic disease. The effect of inoculation dose of Sarcocystis neurona sporocysts on the development of clinical neurologic disease in horses was investigated. Twenty-four seronegative weanling horses were subjected to the natural stress of transport and then randomly assigned to 6 treatment groups of 4 horses each. Horses were then immediately inoculated with either 10(2), 10(3), 10(4), 10(5), or 10(6) S. neurona sporocysts or placebo using nasogastric tube and housed indoors. Weekly neurologic examinations were performed by a blinded observer. Blood was collected weekly for antibody determination by Western ...
Cloning and molecular characterization of an immunogenic LigA protein of Leptospira interrogans. A clone expressing a novel immunoreactive leptospiral immunoglobulin-like protein A of 130 kDa (LigA) from Leptospira interrogans serovar pomona type kennewicki was isolated by screening a genomic DNA library with serum from a mare that had recently aborted due to leptospiral infection. LigA is encoded by an open reading frame of 3,675 bp, and the deduced amino acid sequence consists of a series of 90-amino-acid tandem repeats. A search of the NCBI database found that homology of the LigA repeat region was limited to an immunoglobulin-like domain of the bacterial intimin binding protein of Esc...
Evaluation of 5 serologic assays to detect Rhodococcus equi pneumonia in foals. To determine the sensitivity and specificity of 5 serologic assays used to diagnose Rhodococcus equi pneumonia in foals and to determine whether any of the assays could be used to identify affected foals prior to the onset of clinical signs or to differentiate between affected and unaffected foals when clinical signs first become apparent. Methods: Nested case-control study. Methods: 26 foals. Methods: Serum samples were obtained from all foals at 2, 4, and 6 or 7 weeks of age. Additional samples were obtained from affected foals at the time of diagnosis of R equi pneumonia and from age-matche...
Diagnosis of equine piroplasmosis in Xinjiang province of China by the enzyme-linked immunosorbent assays using recombinant antigens. The prevalence of equine piroplasmosis in Xinjiang province, China, was examined by enzyme-linked immunosorbent assays (ELISAs). A total of 70 serum samples were taken from horses pastured on three farms in western Xinjiang, and examined for diagnosis of equine Babesia equi (B. equi) infection and B. caballi infection by ELISAs using recombinant equi merozoite antigen 1 (EMA-1) and recombinant P48 antigen, respectively. Of the 70 samples, 28 (40.0%) and 17 (24.3%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 11 (15.7%) samples were positive f...
The use of chosen serological diagnostic methods in Lyme disease in horses. Part II. Western blot. In this investigation the Western blot test was treated as a method verifying results of the IFA, commercial ELISA and standardized ELISA tests (described in Part I). The verifying investigations were performed on 82 serum samples, which in the commercial ELISA were positive in 36 cases, dubious in 31 cases and negative in 15 cases as well as on 5 serum samples obtained from horses infected with Leptospira spp., which in the ELISA commercial were dubious (total of 87 sera samples). The antigens, against which the immunological response in horses was directed, were also established. The Milenia...
The use of chosen serological diagnostic methods in Lyme disease in horses. Part I. Indirect immunofluorescence and enzyme-linked immunosorbent assay (ELISA). The investigations aimed to establish the reliability of the chosen serological tests designed for the diagnosis of Lyme borreliosis in horses. The investigations were carried out in five Horse Breeding Centres (OHK). Statistical analysis methods were used to determine sample size for particular centres: Krasne (Kr)--49, Łack (Ł)--21, Walewice (W)--111, BogusŁawice (B)--17, Kozienice (K)--61. The experimental material comprised the chosen horses from which blood samples were collected in order to obtain sera. The test used for indirect immunofluorescence assay (IFA No 75941, Bio-Mérieux) i...
The classification of seven serotypes of equine encephalosis virus and the prevalence of homologous antibody in horses in South Africa. Selected isolates of equine encephalosis virus were shown to have comparable viral protein profiles and to represent seven distinct serotypes, based on cross-neutralization tests. Serotype-specific virus-neutralizing antibody in serum samples from horses confirmed the widespread occurrence of infection. The distribution and prevalence of individual serotypes however, varied considerably. Localised foci with an increased seasonal seroconversion in groups of horses to a specific serotype and the detection of an ongoing low level of infection from other serotypes within the population, confirmed ...
Seroepidemiologic studies on Babesia caballi and Babesia equi infections in Japan. Antibodies to Babesia caballi and Babesia equi were examined on a total of 2,019 horse serum samples that had been collected in 1971-1973 by the National Institute of Animal Health by enzyme-linked immunosorbent assay (ELISA) using recombinant proteins and by Western-blot analysis. Based on the criterion for positivity by ELISA, 5.4% (109/2,019) and 2.2% (44/2,019) had antibodies against B. caballi and B. equi, respectively. The ELISA-positive sera were further examined by Western blot; 30/109 for B. caballi and 2/ 44 for B. equi were positive for native B. caballi or B. equi, but none of them...
Importance of M-protein C terminus as substrate antigen for serodetection of equine arteritis virus infection. Equine arteritis virus (EAV), an enveloped positive-stranded RNA virus, is the prototype of the arterivirus group. In a previous paper (A. Kheyar, S. Martin, G. St.-Laurent, P. J. Timoney, W. H. McCollum, and D. Archambault, Clin. Diagn. Lab. Immunol. 4:648-652, 1997), we have shown that the unglycosylated membrane (M) protein, which is composed of 162 amino acids (aa), is a major target of equine antibody to EAV. In order to determine the antigenic regions of the M protein, the cDNA encoding the M protein of EAV was inserted into the procaryotic expression vector pGEX-4T-1 to produce recombin...
Diagnosis of equine piroplasmosis in Brazil by serodiagnostic methods with recombinant antigens. Serum samples from horses in the States of Sao Paulo and Mato Grosso do Sul, Brazil were examined for diagnosis of equine piroplasmosis by both the latex agglutination test (LAT) and enzyme-linked immunosorbent assay (ELISA) with recombinant antigens. Of the 47 samples analyzed, 38 (81%) and 42 (90%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 35 (75%) samples were positive for both B. equi and B. caballi infections. These results indicate that equine piroplasmosis is widespread and therefore a cause for serious concern in the States of Sao ...
Serum and vitreous humor antibody titers in and isolation of Leptospira interrogans from horses with recurrent uveitis. To measure antibody titers against Leptospira interrogans in serum and vitreous humor and determine the prevalence of L interrogans in vitreous humor of horses with recurrent uveitis. Methods: Cross-sectional study. Methods: 242 horses (270 eyes) with recurrent uveitis undergoing vitrectomy and 39 control horses (54 eyes) without any history or clinical signs of recurrent uveitis undergoing euthanasia or enucleation for unrelated reasons. Methods: Serum and vitreous humor were tested for antibodies against 13 serovars of L interrogans. Vitreous humor was submitted for leptospiral culture; isol...
Immunodiagnosis of Trichinella infection in the horse. From 1998 to 2000, 5,267 horse sera were collected from several Trichinella regions in Romania. Sera were initially screened in laboratories in Romania, Serbia and Italy with an ELISA and a Western blot (Wb) using an excretory/secretory (ES) antigen and several conjugates (protein A, protein G, and sheep or goat anti-horse). Differences in serology results were obtained among the different conjugates and also between ELISA and Wb. Depending on the test used, specific antibodies were found at a prevalence rate of 3-6% of horses. Serum samples classified as positive were tested again by ELISA us...
Validation of a competitive enzyme-linked immunosorbent assay for diagnosing Babesia equi infections of Moroccan origin and its use in determining the seroprevalence of B. equi in Morocco. 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...
Detection of antibodies to the nonstructural protein (NS1) of influenza A virus allows distinction between vaccinated and infected horses. Antibodies to the nonstructural protein (NS1) of A/equine/Miami/1/63 (H3N8) influenza virus were detected exclusively in the sera of mice experimentally infected with A/Aichi/2/68 (H3N2) and horses infected with A/equine/Kentucky/1/81 (H3N8) or A/equine/La Plata/1/93 (H3N8), but not in those of the animals immunized with the inactivated viruses, by enzyme-linked immunosorbent assay (ELISA) using a recombinant NS1 as antigen. The results indicate that the present method is useful for serological diagnosis to distinguish horses infected with equine H3 influenza viruses from those immunized with ...
Development of an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay for detection of equine and swine IgM antibodies to vesicular stomatitis virus. An immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (MC-ELISA) was developed for the detection of primary infection of vesicular stomatitis virus (VSV) in equine and swine sera. The test was based on the use of biotinylated sheep antibodies against equine or swine IgM molecules bound to a streptavidin-coated ELISA plate. The captured IgM antibodies were detected by application of antigens prepared from the New Jersey and the Indiana VSV serotypes (VSV-NJ and VSV-IN, respectively) and mouse polyclonal antibodies against VSV-NJ and VSV-IN. The MC-ELISA was compared to a competiti...
Detection of antibodies to Babesia equi in horses by a latex agglutination test using recombinant EMA-1. A latex agglutination test (LAT) using recombinant equi merozoite antigen 1 (EMA-1) for the detection of antibodies to Babesia equi was developed. The LAT was able to differentiate very clearly between sera from B. equi-infected horses and sera from Babesia caballi-infected horses or from normal horses. The LAT results were identical to those of a previously developed enzyme-linked immunosorbent assay. These results indicate that LAT using recombinant EMA-1 might be very useful as a routine screening method for the diagnosis of B. equi infection.
B-Cell epitope mapping of the VapA protein of Rhodococcus equi: implications for early detection of R. equi disease in foals. Linear B-cell epitopes of the Rhodococcus equi virulence-associated protein (VapA) were mapped using a synthetic peptide bank in this study. The peptides were screened in an enzyme-linked immunosorbent assay (ELISA) with a total of 70 sera from foals with current R. equi disease (51 sera), as well as from foals that had either recovered from R. equi infection 10 months previously (3 sera) or that had no known history of R. equi disease (16 sera). An epitope with the sequence NLQKDEPNGRA was identified and was universally recognized by all 51 sera from foals with R. equi disease and was not rec...
Neurological disease associated with EHV-1-infection in a riding school: clinical and virological characteristics. An outbreak of neurological disease caused by EHV-1 infection is described with emphasis on diagnosis and prognosis for recumbent horses. In April 1995, an outbreak of the neurological form of Equine herpesvirus type 1 (EHV-1) occurred in a well-managed riding school with 41 horses: 34 horses showed a temperature spike and 20 some degree of neurological signs, of which 10 were nursed intensively in the indoor arena of the riding school for 3 to 20 days, 8 having to be maintained in slings for 2-18 days, while 9 needed bladder catheterisation b.i.d. for 2-16 days. Within the first 3 days, one h...
An enzyme-linked immunosorbent assay for the convenient serodiagnosis of contagious equine metritis in mares. An enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of contagious equine metritis (CEM), a sexually transmitted disease caused by Taylorella equigenitalis. Antigen preparation was simple, and antigens derived from both classical and atypical forms of T. equigenitalis enabled detection of antibody responses elicted in horses experimentally exposed to either form of the bacterium. Sera serially obtained from these horses from 0 to 63 days postexposure were tested by the traditional complement fixation test (CFT) for CEM and with the ELISA, using both antigens separat...
Interpretation of the detection of Sarcocystis neurona antibodies in the serum of young horses. Horses that are exposed to Sarcocystis neurona, a causative agent of equine protozoal myeloencephalitis, produce antibodies that are detectable in serum by western blot (WB). A positive test is indicative of exposure to the organism. Positive tests in young horses can be complicated by the presence of maternal antibodies. Passive transfer of maternal antibodies to S. neurona from seropositive mares to their foals was evaluated. Foals were sampled at birth (presuckle), at 24h of age (postsuckle), and at monthly intervals. All foals sampled before suckling were seronegative. Thirty-three foals f...
Direct agglutination test for the detection of antibodies to Sarcocystis neurona in experimentally infected animals. Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in the Americas. The apicomplexan protozoan most commonly associated with EPM is Sarcocystis neurona. A direct agglutination test (SAT) was developed to detect antibodies to S. neurona in experimentally infected animals. Merozoites of the SN6 strain of S. neurona collected from cell culture were used as antigen and 2-mercaptoethanol was added to the antigen suspension to destroy IgM antibodies when mixed with test sera. Mice fed sporocysts of S. speeri or S. falcatula-like sporocysts from opossums did not sero...
Evaluation of a polyvalent enzyme-linked immunosorbent assay incorporating a recombinant p44 antigen for diagnosis of granulocytic ehrlichiosis in dogs and horses. To develop and evaluate a polyvalent ELISA incorporating a highly specific recombinant antigen (p44) for diagnosis of granulocytic ehrlichiosis in dogs and horses. Methods: 32 dogs and 43 horses. Methods: Results of the ELISA were compared with results of indirect fluorescent antibody (IFA) staining and western immunoblotting incorporating whole-cell antigen. Results: For the canine and equine samples, percentages of samples with positive IFA staining, western immunoblotting, and ELISA results were similar. For 29 (91 %) canine samples and 30 (70%) equine samples, results of IFA staining, west...