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Topic:Antibodies

Antibodies in horses are specialized proteins produced by the immune system in response to foreign substances, known as antigens. These substances can include pathogens such as bacteria, viruses, and parasites. Antibodies function by recognizing and binding to specific antigens, thereby neutralizing them or marking them for destruction by other immune cells. In equine health, antibodies are integral to both natural immune responses and those induced by vaccinations. The study of antibodies in horses encompasses their production, diversity, and role in disease resistance and management. This page gathers peer-reviewed research studies and scholarly articles that explore the generation, function, and implications of antibodies in equine immunology and disease control.
Absence of evidence for bornavirus infection in schizophrenia, bipolar disorder and major depressive disorder.
Molecular psychiatry    January 31, 2012   Volume 17, Issue 5 486-493 doi: 10.1038/mp.2011.179
Hornig M, Briese T, Licinio J, Khabbaz RF, Altshuler LL, Potkin SG, Schwemmle M, Siemetzki U, Mintz J, Honkavuori K, Kraemer HC, Egan MF, Whybrow PC....In 1983, reports of antibodies in subjects with major depressive disorder (MDD) to an as-yet uncharacterized infectious agent associated with meningoencephalitis in horses and sheep led to molecular cloning of the genome of a novel, negative-stranded neurotropic virus, Borna disease virus (BDV). This advance has enabled the development of new diagnostic assays, including in situ hybridization, PCR and serology based on recombinant proteins. Since these assays were first implemented in 1990, more than 80 studies have reported an association between BDV and a wide range of human illnesses that i...
Vitreal IgM autoantibodies target neurofilament medium in a spontaneous model of autoimmune uveitis.
Investigative ophthalmology & visual science    January 25, 2012   Volume 53, Issue 1 294-300 doi: 10.1167/iovs.11-8734
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...
Analysis of antibody responses by commercial western blot assay in horses with alveolar echinococcosis.
The Journal of veterinary medical science    January 20, 2012   Volume 74, Issue 6 813-815 doi: 10.1292/jvms.11-0552
Ueno M, Kuroda N, Yahagi K, Ohtaki T, Kawanaka M.Commercial western blot (WB) assay was used to detect serum antibodies specific to Echinococcus multilocularis in 23 horses in which infection was confirmed by postmortem inspection at a slaughterhouse. Livers contained from 1 to >20 nodular lesions; foci diameter ranged from 1 to 25 mm. Antibody tests of serum from all 23 animals were negative for antigen bands at 7, 16, 18, and 26-28 kDa, which show specificity in the serum of human patients. However, sera from two infected horses with the largest nodules (diameter, 25 mm) showed positive response to one of the 22-kDa and 30-kDa antigen band...
Clinical and virological outcome of an infection with the Belgian equine arteritis virus strain 08P178.
Veterinary microbiology    January 18, 2012   Volume 157, Issue 3-4 333-344 doi: 10.1016/j.vetmic.2012.01.014
Vairo S, Vandekerckhove A, Steukers L, Glorieux S, Van den Broeck W, Nauwynck H.Equine viral arteritis (EVA) is an infectious disease with variable clinical outcome. Outbreaks, causing important economic losses, are becoming more frequent. Currently, there is a shortage of pathogenesis studies performed with European strains. In the present study, eight seronegative ponies were experimentally inoculated with the Belgian strain of equine arteritis virus (EAV) 08P178 (EU-1 clade) and monitored daily for clinical signs of EVA. Nasopharyngeal swabs, ocular swabs, bronchoalveolar cells and blood were collected for virological and serological testing. Two ponies were euthanized...
Maintaining health in foals: the role of colostrum constituents.
The Veterinary record    January 17, 2012   Volume 170, Issue 2 49-50 doi: 10.1136/vr.e187
Davis E.No abstract available
Antibodies to a novel leptospiral protein, LruC, in the eye fluids and sera of horses with Leptospira-associated uveitis.
Clinical and vaccine immunology : CVI    January 11, 2012   Volume 19, Issue 3 452-456 doi: 10.1128/CVI.05524-11
Verma A, Matsunaga J, Artiushin S, Pinne M, Houwers DJ, Haake DA, Stevenson B, Timoney JF.Screening of an expression library of Leptospira interrogans with eye fluids from uveitic horses resulted in identification of a novel protein, LruC. LruC is located in the inner leaflet of the leptospiral outer membrane, and an lruC gene was detected in all tested pathogenic L. interrogans strains. LruC-specific antibody levels were significantly higher in eye fluids and sera of uveitic horses than healthy horses. These findings suggest that LruC may play a role in equine leptospiral uveitis.
Surveillance for antibodies to West Nile virus in Ireland.
The Veterinary record    January 11, 2012   Volume 170, Issue 7 180 doi: 10.1136/vr.100333
Raleigh PJ, Sammin DJ, Connell J, Markey BK, O'Connor M.No abstract available
A diagnostic algorithm to serologically differentiate West Nile virus from Japanese encephalitis virus infections and its validation in field surveillance of poultry and horses.
Vector borne and zoonotic diseases (Larchmont, N.Y.)    January 4, 2012   Volume 12, Issue 5 372-379 doi: 10.1089/vbz.2011.0709
Yeh JY, Lee JH, Park JY, Seo HJ, Moon JS, Cho IS, Kim HP, Yang YJ, Ahn KM, Kyung SG, Choi IS, Lee JB.The detection of West Nile virus (WNV) in areas endemic for Japanese encephalitis virus (JEV) is complicated by the extensive serological cross-reactivity between the two viruses. A testing algorithm was developed and employed for the detection of anti-WNV antibody in areas endemic for JEV. Using this differentiation algorithm, a serological survey of poultry (2004 through 2009) and horses (2007 through 2009) was performed. Among 2681 poultry sera, 125 samples were interpreted as being positive for antibodies against JEV, and 14 were suspected to be positive for antibodies against undetermined...
Development and laboratory evaluation of two lateral flow devices for the detection of vesicular stomatitis virus in clinical samples.
Journal of virological methods    December 29, 2011   Volume 180, Issue 1-2 96-100 doi: 10.1016/j.jviromet.2011.12.010
Ferris NP, Clavijo A, Yang M, Velazquez-Salinas L, Nordengrahn A, Hutchings GH, Kristersson T, Merza M.Two lateral flow devices (LFD) for the detection of vesicular stomatitis (VS) virus (VSV), types Indiana (VSV-IND) and New Jersey (VSV-NJ) were developed using monoclonal antibodies C1 and F25VSVNJ-45 to the respective VSV serotypes. The performance of the LFDs was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture passage derived supernatants of VSV. The collection of test samples included 105 positive for VSV-IND (92 vesicular epithelial suspensions and 13 cell culture antigens; encompassing 93 samples of subtype 1 [VSV-IND-1], 9 of subtype 2 [VSV-IND-2] and 3...
Immunological correlates of vaccination and infection for equine herpesvirus 1.
Clinical and vaccine immunology : CVI    December 28, 2011   Volume 19, Issue 2 235-241 doi: 10.1128/CVI.05522-11
Goodman LB, Wimer C, Dubovi EJ, Gold C, Wagner B.Equine herpesvirus 1 (EHV-1) induces a variety of disease manifestations, including respiratory disease, abortions, and myeloencephalopathy. Several vaccines are commercially available but could not previously be distinguished by serologic testing from infection with EHV-1 (or the closely related EHV-4). Currently available vaccines are not reliably protective against the severe manifestations of the disease, including fatal myeloencephalopathy. We determined immunological parameters that can differentiate vaccinated from previously infected animals by comparing humoral and cellular EHV-1-spec...
Animal diseases caused by orbiviruses, Algeria.
Emerging infectious diseases    December 17, 2011   Volume 17, Issue 12 2325-2327 doi: 10.3201/eid1712.110928
Madani H, Casal J, Alba A, Allepuz A, Cêtre-Sossah C, Hafsi L, Kount-Chareb H, Bouayed-Chaouach N, Saadaoui H, Napp S.Antibodies against bluetongue virus were detected in cattle, sheep, goats, and camels in Algeria in 2008. Antibodies against epizootic hemorrhagic disease virus were detected in cattle, but antibodies against African horse sickness virus were not detected in horses and mules. Epizootic hemorrhagic disease in northern Africa poses a major risk for the European Union.
Seroprevalence of Toxoplasma gondii in domestic and wild animals from the Fernando de Noronha, Brazil.
The Journal of parasitology    December 12, 2011   Volume 98, Issue 3 679-680 doi: 10.1645/GE-2910.1
Costa DG, Marvulo MF, Silva JS, Santana SC, Magalhães FJ, Filho CD, Ribeiro VO, Alves LC, Mota RA, Dubey JP, Silva JC.Fernando de Noronha is an archipelago of 21 islands and islets in the Atlantic Ocean, state of Pernambuco, Brazil, which has a varied biodiversity including alien species or sinantropic animals. The objective here was to determine the seroprevalence of Toxoplasma gondii in domestic and wild animals from Fernando de Noronha archipelago, Brazil. Between July 2007 and May 2010, blood samples were collected from 764 animals (533 domestic and 231 wild animals). Sera were tested by the indirect fluorescence antibody test (IFAT) or the modified agglutination test (MAT), or by both. Antibodies to T. g...
Adaptation and validation of a bacteria-specific enzyme-linked immunosorbent assay for determination of farm-specific Lawsonia intracellularis seroprevalence in central Kentucky Thoroughbreds.
Equine veterinary journal. Supplement    December 7, 2011   Issue 40 25-31 doi: 10.1111/j.2042-3306.2011.00487.x
Page AE, Stills HF, Chander Y, Gebhart CJ, Horohov DW.Lawsonia intracellularis is the causative agent of equine proliferative enteropathy (EPE), a disease for which no large-scale seroprevalence studies have been conducted. Objective: To validate and use an equine-specific enzyme-linked immunosorbent assay (ELISA) for L. intracellularis to determine the seroprevalence of L. intracellularis on numerous farms. Methods: An ELISA, in which purified antigen was used, was adapted from previous work in swine. A total of 337 Thoroughbreds from 25 central Kentucky farms were enrolled and monthly serum samples collected from August 2010 to January/February...
Serosurveillance for Japanese encephalitis virus infection among equines in India.
Journal of veterinary science    November 30, 2011   Volume 12, Issue 4 341-345 doi: 10.4142/jvs.2011.12.4.341
Gulati BR, Singha H, Singh BK, Virmani N, Khurana SK, Singh RK.The seroprevalence of Japanese encephalitis virus (JEV) among equines was evaluated from January 2006 to December 2009 in 13 different states of India by hemagglutination inhibition (HI) test and virus neutralization test (VNT). Antibodies against JEV were detected in 327 out of 3,286 (10%) equines with a maximum prevalence reported in the state of Manipur (91.7%) followed by Gujarat (18.5%), Madhya Pradesh (14.4%), and Uttar Pradesh (11.6%). Evidence of JEV infection was observed in equines in Indore (Madhya Pradesh) where a 4-fold or higher rise in antibody titer was observed in 21 out of 34...
Immunohistochemical analysis of cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA) in murine, canine, bovine and equine cerebrospinal tissues.
The Journal of veterinary medical science    November 29, 2011   Volume 74, Issue 4 523-526 doi: 10.1292/jvms.11-0494
Tokunaga S, Fujiki M, Yabuki A, Misumi K.Cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA), which appears abundantly in hypertrophic cartilage at the stage of endochondral ossification, is also detected in cerebrospinal fluid (CSF) following spinal cord injury. In this study, the localization of the CD-RAP/MIA molecule in normal tissues of the spine and brain obtained from mice, rats, dogs, cattle and horses was examined using immunohistochemistry with a specific antibody. The positive signals of CD-RAP/MIA were found at nerve cells in the spinal cords of all species and were especially str...
ISCOM-matrix-based equine influenza (EIV) vaccine stimulates cell-mediated immunity in the horse.
Veterinary immunology and immunopathology    November 26, 2011   Volume 145, Issue 1-2 516-521 doi: 10.1016/j.vetimm.2011.11.019
Paillot R, Prowse L.The humoral immune response induced by ISCOM-matrix (Immuno Stimulating COMplex-Matrix)-adjuvanted equine influenza virus (EIV) vaccine is well documented in horses. ISCOM-matrix adjuvanted vaccines against human influenza are strong inducers of cell-mediated immunity (CMI), including T cell proliferation and virus-specific cytotoxic T cell. In the horse, the CMI response to equine influenza vaccination is less well characterised. An ISCOM-based vaccine has been shown to induce interferon gamma (IFN-γ) synthesis, a CMI marker, in the horse, but this has not been shown for the ISCOM-matrix vac...
Seroprevalence of Toxoplasma gondii infection among horses in Tunisia.
Parasites & vectors    November 22, 2011   Volume 4 218 doi: 10.1186/1756-3305-4-218
Boughattas S, Bergaoui R, Essid R, Aoun K, Bouratbine A.The present study was conducted to investigate the serological survey of Toxoplasma antibodies in local.horses from three major regions: a neighbourhood of a city in the North (Sidi Thabet), a neighbourhood of a city on the coast (Monastir) and a neighbourhood of a city in the middle (Battan) of Tunisia (North of Africa). Methods: A total of 158 serum samples were obtained from clinically healthy horses which consisted of 111 (32 female, 79 male) 2-10 years old and 47 (11 female, 36 male) older than 10 years. All of the horses were tested for antibodies to T. gondii using the Modified Agglutin...
Toxoplasma gondii in wild and domestic animals from New Caledonia.
Parasite (Paris, France)    November 18, 2011   Volume 18, Issue 4 345-348 doi: 10.1051/parasite/2011184345
Roqueplo C, Halos L, Cabre O, Davoust B.Samples (serum or meat juice) collected from 205 animals in New Caledonia in April 2009 were tested for antibodies against Toxoplasma gondii by ELISA using the multi-species ID Screen® Toxoplasmosis Indirect kit (IDVET, Montpellier). Antibodies to T. gondii were detected in 2% (1/49) of the pigs, in 3.3% (1/30) of the cattle, in 13.8% (4/29) of Rusa deers, in 16% (4/25) of the horses, in 32.8% (21/64) of the dogs, and in 50% (4/8) of cats. Statistically, no significant difference was observed between T. gondii seroprevalence and age or sex. No survey on the prevalence of T. gondii in anima...
A prospective longitudinal study of naturally infected horses to evaluate the performance characteristics of rapid diagnostic tests for equine influenza virus.
Veterinary microbiology    November 6, 2011   Volume 156, Issue 3-4 246-255 doi: 10.1016/j.vetmic.2011.10.031
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...
Protective effects of passively transferred merozoite-specific antibodies against Theileria equi in horses with severe combined immunodeficiency.
Clinical and vaccine immunology : CVI    October 28, 2011   Volume 19, Issue 1 100-104 doi: 10.1128/CVI.05301-11
Mealey RH, Kappmeyer LS, Ueti MW, Wagner B, Knowles DP.Theileria equi immune plasma was infused into young horses (foals) with severe combined immunodeficiency. Although all foals became infected following intravenous challenge with homologous T. equi merozoite stabilate, delayed time to peak parasitemia occurred. Protective effects were associated with a predominance of passively transferred merozoite-specific IgG3.
Effects of opsonization of Rhodococcus equi on bacterial viability and phagocyte activation.
American journal of veterinary research    October 26, 2011   Volume 72, Issue 11 1465-1475 doi: 10.2460/ajvr.72.11.1465
Dawson DR, Nydam DV, Price CT, Graham JE, Cynamon MH, Divers TJ, Felippe MJ.To investigate the effect of opsonization of Rhodococcus equi with R. equi-specific antibodies in plasma on bacterial viability and phagocyte activation in a cell culture model of infection. Methods: Neutrophils and monocyte-derived macrophages from 6 healthy 1-week-old foals and 1 adult horse. Methods: Foal and adult horse phagocytes were incubated with either opsonized or nonopsonized bacteria. Opsonization was achieved by use of plasma containing high or low concentrations of R. equi-specific antibodies. Phagocyte oxidative burst activity was measured by use of flow cytometry, and macrophag...
Inhibition of cytochrome P450 enzymes involved in ketamine metabolism by use of liver microsomes and specific cytochrome P450 enzymes from horses, dogs, and humans.
American journal of veterinary research    October 26, 2011   Volume 72, Issue 11 1505-1513 doi: 10.2460/ajvr.72.11.1505
Mössner LD, Schmitz A, Theurillat R, Thormann W, Mevissen M.To identify and characterize cytochrome P450 enzymes (CYPs) responsible for the metabolism of racemic ketamine in 3 mammalian species in vitro by use of chemical inhibitors and antibodies. Methods: Human, canine, and equine liver microsomes and human single CYP3A4 and CYP2C9 and their canine orthologs. Methods: Chemical inhibitors selective for human CYP enzymes and anti-CYP antibodies were incubated with racemic ketamine and liver microsomes or specific CYPs. Ketamine N-demethylation to norketamine was determined via enantioselective capillary electrophoresis. Results: The general CYP inhibit...
Exposure to Sarcocystis spp. in horses from Spain determined by Western blot analysis using Sarcocystis neurona merozoites as heterologous antigen.
Veterinary parasitology    October 7, 2011   Volume 185, Issue 2-4 301-304 doi: 10.1016/j.vetpar.2011.09.042
Arias M, Yeargan M, Francisco I, Dangoudoubiyam S, Becerra P, Francisco R, Sánchez-Andrade R, Paz-Silva A, Howe DK.Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocyst...
Seroprevalence of Leptospira spp. in clinically healthy horses in Switzerland.
Schweizer Archiv fur Tierheilkunde    October 6, 2011   Volume 153, Issue 10 449-456 doi: 10.1024/0036-7281/a000247
Blatti S, Overesch G, Gerber V, Frey J, Hüssy D.A retrospective, cross-sectional study was conducted to determine the leptospiral seroprevalence in clinically healthy horses in Switzerland. A representative sample of 615 horse sera was examined by microscopic agglutination test for the presence of antibodies against 15 Leptospira spp. serovars. In total, 58.5 % (n = 360) of the horses were positive for one or more of the antigens analysed, with 20.3 % of them showing titres >= 400. The most prevalent serovar was Pyrogenes (22.6 %), followed by serovars Canicola (22.1 %) and Australis (19.2 %). Older horses, mares, ponies and ani...
A comparison of antibody responses to commercial equine influenza vaccines following primary vaccination of Thoroughbred weanlings–a randomised blind study.
Vaccine    October 5, 2011   Volume 29, Issue 49 9214-9223 doi: 10.1016/j.vaccine.2011.09.101
Gildea S, Arkins S, Walsh C, Cullinane A.Many racing authorities, sales companies and equestrian bodies have mandatory vaccination policies for equine influenza (EI). The consequences of lack of vaccine efficacy include clinical disease, disruption to training programmes, the cancellation of equestrian events and the introduction of virus to susceptible populations. The correlation between antibody against the virus haemagglutinin and protection against influenza has been well established. The objective of this study was to compare the antibody responses of 66 unvaccinated Thoroughbred weanlings on four different stud farms, followin...
Equine ELISPOT assay to study secretion of IFNγ and IL-4 from peripheral blood mononuclear cells.
Methods in molecular biology (Clifton, N.J.)    October 1, 2011   Volume 792 39-45 doi: 10.1007/978-1-61779-325-7_3
Hagen J, Hartnett C, Houchins JP, Giguère S, Kalyuzhny AE.Human and mouse immune system cells are the most frequently used specimens in ELISPOT assays. In an effect to expand the application of ELISPOT assay to other species, we developed matched antibody pairs for ready-to-use kits designed for studying the frequency of equine IFNγ- and IL-4-secreting peripheral blood mononuclear cells (PBMCs). Equine PBMCs were stimulated with either concanavalin A (Con A) or calcium ionomycin mixed with phorbol 12-myristate 13-acetate (CaI + PMA). We found that Con A, in general, had a more profound stimulating effect than CaI + PMA on IL-4 secretion, whereas bot...
Safety and immunogenicity of BPV-1 L1 virus-like particles in a dose-escalation vaccination trial in horses.
Equine veterinary journal    September 6, 2011   Volume 44, Issue 1 107-111 doi: 10.1111/j.2042-3306.2011.00390.x
Hainisch EK, Brandt S, Shafti-Keramat S, Van den Hoven R, Kirnbauer R.Infection with bovine papillomaviruses types 1 and 2 (BPV-1, BPV-2) can lead to the development of therapy-resistant skin tumours termed sarcoids and possibly other skin diseases in equids. Although sarcoids seriously compromise the welfare of affected animals and cause considerable economic losses, no prophylactic vaccine is available to prevent this common disease. In several animal species and man, immunisation with papillomavirus-like particles (VLP) has been shown to protect efficiently from papillomaviral infection. Objective: BPV-1 L1 VLPs may constitute a safe and highly immunogenic va...
Antimicrobial peptides and proteins of the horse–insights into a well-armed organism.
Veterinary research    September 2, 2011   Volume 42, Issue 1 98 doi: 10.1186/1297-9716-42-98
Bruhn O, Grötzinger J, Cascorbi I, Jung S.Antimicrobial peptides play a pivotal role as key effectors of the innate immune system in plants and animals and act as endogenous antibiotics. The molecules exhibit an antimicrobial activity against bacteria, viruses, and eukaryotic pathogens with different specificities and potencies depending on the structure and amino-acid composition of the peptides. Several antimicrobial peptides were comprehensively investigated in the last three decades and some molecules with remarkable antimicrobial properties have reached the third phase of clinical studies. Next to the peptides themselves, numerou...
[Serological study carried out in Cuban localities where confirmed western Nile virus infection is present].
Revista cubana de medicina tropical    September 1, 2011   Volume 63, Issue 3 227-230 
first infected cases caused by West Nile virus were reported in Cuba in 2004. Objective: to monitor and learn about the prevalence of the West Nile virus in those areas with confirmed cases. Methods: the study was conducted in Jatibonico municipality and in the city of sancti Spiritus. A total number of 14 persons, 8 horses and 41 birds were researched to detect antibodies to flavivirus and specific antibodies to West Nile virus. Results: the presence of specific antibodies to West Nile virus was confirmed in 4 samples of sera from birds and in 4 from horses. One person was confirmed as one ca...
Mapping B-cell epitopes in equine rhinitis B viruses and identification of a neutralising site in the VP1 C-terminus.
Veterinary microbiology    August 30, 2011   Volume 155, Issue 2-4 128-136 doi: 10.1016/j.vetmic.2011.08.022
Horsington JJ, Gilkerson JR, Hartley CA.Erbovirus is a genus of the family Picornaviridae and equine rhinitis B virus (ERBV) is the sole species. Erboviruses infect horses causing acute respiratory disease and sub-clinical and persistent infections. Despite the high seroprevalence and worldwide distribution of these viruses, the pathogenesis and antigenic structure of the three ERBV serotypes (ERBV1, 2 and 3) is poorly understood. To characterise linear epitopes on ERBV structural proteins, a set of fusion proteins were expressed in Escherichia coli. These proteins were tested in Western blot and ELISA and reactive proteins were als...
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