Serodiagnosis of equine infectious anemia by agar gel immunodiffusion and ELISA using a recombinant p26 viral protein expressed in Escherichia coli as antigen

Piza, Adriana S. Toledo; Pereira, Alessandra Rael; Terreran, Maria Thereza; Mozzer, O. D.; Tanuri, Amílcar; Brandão, Paulo Eduardo; Richtzenhain, Leonardo José

doi:10.1016/j.prevetmed.2006.10.009

Cited by 15 publications

(7 citation statements)

References 14 publications

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“…This is the first report of optimizing a diagnostic ELISA and AGID in large number of field serum samples (n = 7,150 in ELISA, and n = 1,200 in AGID) for diagnosis of EIA employing gene synthesis technology. Diagnostic sensitivity and specificity rp26-ELISA and rp26-AGID obtained in the present study are in complete agreements with previous findings [21,22,28,29]. Our results suggest that the assays can be used for a large-scale screening test to monitor the prevalence of EIA and assess the risk factors.…”

Section: Discussionsupporting

confidence: 92%

“…In an effort to develop a more rapid and sensitive diagnostic capability, several attempts have been made on development of enzyme-linked immunosorbent assays (ELISAs) based on p26 protein purified from EIAV strain [28,29,32], recombinant proteins [1,2,15,22,25,33] and peptides [27,31]. The EIAV p26 protein encoded by gag gene is one of the highly conserved structural proteins among the viral isolates even from very different geographical locations [4,5,26].…”

Section: Discussionmentioning

confidence: 99%

“…However, recent emergence of EIAV infection in Asian continent like, Japan [17], Mongolia [20], China, Thailand, Uzbekistan, Philippines, and Malaysia during [2008][2009][2010][2011][2012] [http://www.oie.int/wahis_2/public/wahid.php/Wahidhome] and detection of EIAV infection in 2010 and 2012 in India [16] after a long gap underlines the importance of continued surveillance to monitor the disease prevalence and identifying the genetic variation among field strains of EIAV. During the last two decades, the detection of EIAV antibodies by a number of ELISAs has been described and compared with 'gold standard' agar gel immuno-diffusion (AGID) test [3,21,22,24,25,28,29,31,32]. The ELISA has been reported to be more sensitive than AGID in detecting acute cases of EIA and helpful in disease eradication programme [10].…”

Section: Introductionmentioning

confidence: 99%

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Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus

et al. 2013

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Equine infectious anemia (EIA)-a retroviral disease caused by equine infectious anemia virus (EIAV)-is a chronic, debilitating disease of horses, mules, and donkeys. EIAV infection has been reported worldwide and is recognized as pathogen of significant economic importance to the horse industry. This disease falls under regulatory control program in many countries including India. Control of EIA is based on identification of inapparent carriers by detection of antibodies to EIAV in serologic tests and ''Stamping Out'' policy. The current internationally accepted test for diagnosis of EIA is the agar gel immune-diffusion test (AGID), which detects antibodies to the major gag gene (p26) product. The objective of this study was to develop recombinant p26 based in-house immunoassays [enzyme linked immunosorbent assays (ELISA), and AGID] for EIA diagnosis. The synthetic p26 gene of EIAV was expressed in Escherichia coli and diagnostic potential of recombinant p26 protein were evaluated in ELISA and AGID on 7,150 and 1,200 equine serum samples, respectively, and compared with commercial standard AGID kit. The relative sensitivity and specificity of the newly developed ELISA were 100 and 98.6 %, respectively. Whereas, relative sensitivity and specificity of the newly developed AGID were in complete agreement in respect to commercial AGID kit. Here, we have reported the validation of an ELISA and AGID on large number of equine serum samples using recombinant p26 protein produced from synthetic gene which does not require handling of pathogenic EIAV. Since the indigenously developed reagents would be economical than commercial diagnostic kit, the rp26 basedimmunoassays could be adopted for the sero-diagnosis and control of EIA in India.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus

et al. 2013

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“…Although the double-antigen ELISA technique used to detect antibodies against EIAV in the current study was not the OIE reference technique (Coggins test), when testing on a larger scale for screening purposes is required within a short period of time, ELISA becomes the test of choice ( Paré and Simard 2004 , Piza and others 2007 ). Some authors have demonstrated a high degree of agreement between specific EIAV genetic material presence and antibodies in equids that are reported as negative by Coggins test but positive by ELISA ( Issel and others 2013 , Scicluna and others 2013 ); ELISA techniques can detect antibodies directed against the p26 EIAV-capsid before those detectable by the Coggins test and overall are considered more sensitive ( Soutullo and others 2001 , Reis and others 2012 ).…”

Section: Discussionmentioning

confidence: 99%

Freedom from equine infectious anaemia virus infection in Spanish Purebred horses

Cruz-López¹,

Fores²,

Ireland

et al. 2015

Veterinary Record Open

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IntroductionNo cases of equine infectious anaemia (EIA) have been reported in Spain since 1983. Factors that could increase the risk of reintroducing equine infectious anaemia virus (EIAV) into Spain include the recent occurrence of the disease in Europe and the absence of compulsory serological testing before importation into Spain.Aims and objectivesGiven the importance of the Spanish Purebred (SP) horse breeding industry in Spain, the aim of this cross-sectional study was to provide evidence of freedom from EIAV in SP stud farms in Central Spain.Materials and methodsSerum samples from 555 SP horses, collected between September 2011 and November 2013, were tested using a commercially available EIAV ELISA with a published sensitivity of 100 per cent.ResultsAll 555 samples were negative for antibody to EIAV, providing evidence of a true EIAV seroprevalence between 0 per cent and 0.53 per cent (95% CIs of the sensitivity and specificity of the ELISA technique used Q10 were 100 per cent and 99.3 per cent, respectively) among the SP breeding population in Central Spain.ConclusionsThese findings should serve to increase confidence when exporting SP horses to other countries.

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“…Agar Gel Immunodiffusion (AGID) is a serological test that the OIE and the Ministry of Agriculture, Livestock and Food Supply (MAPA) consider as the gold standard for diagnosing EIA [4]- [7]. Indeed, the OIE recommends the test for international trade [3].…”

Section: Introductionmentioning

confidence: 99%

Comparison of Two Protocols of Agar Gel Immunodiffusion (AGID) Used to Diagnose of Equine Infectious Anemia (EIA)

Beltrão¹,

Alencar²,

Leite³

et al. 2015

OJVM

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The aim of the present study was to compare two protocols of Agar Gel Immunodiffusion (AGID) that can be used to diagnose Equine Infectious Anemia (EIA). Different volumes of reagents and support for agar gel were used, as recommended by the Ministry of Agriculture, Livestock and Food Supply (MAPA) and the World Organization for Animal Health (OIE). Equidae sera samples (n = 1000) were tested using a national commercial kit and different volumes (i.e. 25 µL-MAPA and 50 µL-OIE) of reagents and standard sera. Antigen was used as recommended by MAPA. The results were assessed by two operators, 48 hours after beginning the test. At the end of the experiment, a Kappa index of 1 was obtained by the operators, for the interpretation of the results and for the different volumes of reagents used. Similar results of agreement (Kappa index = 1) were obtained when using slides or Petri dishes as support for the agar gel. In both cases, an almost perfect agreement was achieved, suggesting that the use of different supports does not affect the results. These findings indicate that both protocols (MAPA and OIE) can be used to diagnose EIA.

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Serodiagnosis of equine infectious anemia by agar gel immunodiffusion and ELISA using a recombinant p26 viral protein expressed in Escherichia coli as antigen

Cited by 15 publications

References 14 publications

Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus

Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus

Freedom from equine infectious anaemia virus infection in Spanish Purebred horses

Comparison of Two Protocols of Agar Gel Immunodiffusion (AGID) Used to Diagnose of Equine Infectious Anemia (EIA)

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