A Review of Techniques for the Serologic Diagnosis of Equine Infectious Anemia

Abstract

History and introduction This paper will review the current serologic tests to Equine infectious anemia (EIA) has been recognized diagnose infections with EIAV, discuss limitations of as an important viral disease of members of the horse the procedures, and suggest improvements based on family since its "filterable virus" etiology was proven early in the twentieth century. 49 Only since the 1970s EIAV and the horse have we been able to diagnose infection with equine infectious anemia virus (EIAV) accurately with the use The EIAV is a member of the lentivirus subfamily of serodiagnostic methods. 6,7 Early attempts to define of retroviruses and shares morphologic, antigenic, and specific diagnostic tools for EIA failed in part because genetic characters with the other lentiviruses. 17 The of a lack of suitable in vivo laboratory animal models major core protein of EIAV is a product of the viral and in vitro techniques for virus propagation, as well gag gene, 45 has a molecular weight of 26,000 daltons

Validation according to OIE criteria of a monoclonal, recombinant p26–based, serologic competitive enzyme-linked immunosorbent assay as screening method in surveillance programs for the detection of Equine infectious anemia virus antibodies

“…Bacterial lysate was then centrifuged at 12,000 × g h for 10 min at 4°C. Both supernatant and pellet were monitored by immunoblot 11,12 for the presence of the p26 recombinant protein. The mAbs employed for the development of the cELISA were supplied by the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (Brescia, Italy), and a brief description of their production and characterization is as follows: mAbs were produced by intramuscular inoculation of BALB/c mice g with the EIAV Wyoming strain (ATCC VR-778) and Freund adjuvant, g followed by 2 inoculations without adjuvant, 1 of which was intraperitoneal.…”

“…11,12 Positive hybridomas were cloned by limiting dilution on a feeder layer of normal mouse spleen cells and injected intraperitoneally in pristane-sensitized mice for production of ascitic fluid; mAbs were purified as previously described. 8 Selection of mAbs was performed by comparing their reciprocal competition for antigen-binding epitopes using ELISA, with p26 as antigen.…”

Comparative study of agar gel immunodiffusion (AGID) protocols for the diagnosis of equine infectious anemia in Brazil

“…These results emphasize the loss of analytical sensitivity of the imported AGID kit when it is used with a lower reagent volume-based procedure. As we mentioned previously, the main limitation of AGID testing is the subjective reading of results COOK, 1993;CULLINANE et al, 2007). Many published studies present equivocal results with weak positive samples due to the difficulty in visualizing the curvature of the precipitation lines in samples with low levels of EIAV antibody (ISSEL; ADAMS, 1982;MCCONNICO et al, 2000).…”

“…The official diagnostic test for EIA in Brazil and in most countries around the world is the AGID (Agar Gel Immunodiffusion) test, also known as the Coggins test (MAPA, 2004;OIE, 2008). This test is simple and highly specific for identifying animals infected with the EIAV, as a positive AGID test correlates with EIAV presence (COGGINS; NORCROSS; NUSBAUM, 1972;COOK, 1993). The basis for the AGID test is the concurrent migration of antigen and antibodies through an agar gel.…”

“…A visible precipitation line is produced when an insoluble antigen-antibody complex becomes visible between the serum and EIAV antigen in the agar gel (COGGINS; NORCROSS; NUSBAUM, 1972). One of the limitations of AGID is that it is considered less sensitive and more subjective in reading than other tests such as ELISA or immunoblotting COOK, 1993;CULLINANE et al, 2007).…”