Purification and characterization of equine infectious anemia virus.
Abstract: EIA virus was purified from equine fetal kidney cell cultures by PEG-precipitation, two sucrose-gradient sedimentations (5-30 per cent) and (25 to 60 per cent) centrifugation, using the immunodiffusion test to follow the procedure. Purified EIA virus had a density (20 degrees C) of 1.162 and a sedimentation constant of S20w=656. electron microscopy revealed a particle of about 100 nm in diameter with a very flexible but usually spherical shape. The dense core may be at various locations inside the membrane bound particle.
Publication Date: 1976-01-01 PubMed ID: 183628DOI: 10.1007/BF01317839Google Scholar: Lookup
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- Journal Article
Summary
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The research paper focuses on purifying and identifying the characteristics of the Equine Infectious Anemia (EIA) virus which causes a highly infectious and potentially deadly disease in horses.
Objective and Methodology
- The researchers carried out the extraction of the EIA virus from cultures of equine fetal kidney cells. This process was followed using a substance called PEG (polyethylene glycol).
- The purified EIA virus was then subjected to two rounds of sucrose-gradient sedimentation processes, which were between 5-30 per cent and 25-60 per cent respectively. This was a step where the virus was separated based on their densities.
- An immunodiffusion test was used to monitor the process. This test helps to identify antigens (foreign substances that induce an immune response in the body) or antibodies (proteins produced by the immune system to neutralize these antigens) in a sample.
Findings
- The findings showed that the purified EIA virus had a specific density of 1.162 (measured at 20 degrees Celsius) and a sedimentation constant of S20w=656. Both of these measures tell us more about the physical characteristics of the virus, and how it behaves in a fluid suspension.
- Electron microscopy was used to visualize the virus. The researchers noticed that the virus particles were roughly 100 nanometers in diameter and had a variable but predominantly spherical shape. Microscopic visualisation provided detailed structural information about the virus.
- They observed that the dense core (the part of the virus containing genetic material and other proteins) could be in various locations within the virus particle. This further enhances our understanding about the structural flexibility of the virus.
Significance
- This research deepens our understanding of the EIA virus, its physical attributes, and behavior. It provides a clearer picture of the virus’ structure, which can aid in the development of treatments or vaccines.
Cite This Article
APA
Matheka HD, Coggins L, Shively JN, Norcross NL.
(1976).
Purification and characterization of equine infectious anemia virus.
Arch Virol, 51(1-2), 107-114.
https://doi.org/10.1007/BF01317839 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Cell Membrane / microbiology
- Centrifugation, Density Gradient
- Chemical Precipitation
- Culture Techniques
- Horses
- Inclusion Bodies, Viral
- Infectious Anemia Virus, Equine / growth & development
- Infectious Anemia Virus, Equine / isolation & purification
- Infectious Anemia Virus, Equine / ultrastructure
- Kidney / embryology
- Leukocytes
References
This article includes 13 references
- Nakajima H, Norcross NL, Coggins L. Demonstration of antigenic identity between purified equine infectious anemia virus and an antigen extracted from infected horse spleen.. Infect Immun 1972 Sep;6(3):416-7.
- Kono Y, Yoshino T. Propagation of equine infectious anemia virus in horse kidney cell cultures.. Natl Inst Anim Health Q (Tokyo) 1974 Winter;14(4):155-62.
- Tajima M, Nakajima H, Ito Y. Electron microscopy of equine infectious anemia virus.. J Virol 1969 Oct;4(4):521-7.
- Coggins L, Norcross NL. Immunodiffusion reaction in equine infectious anemia.. Cornell Vet 1970 Apr;60(2):330-5.
- Nakajima H, Tajima M, Tanaka S, Ushimi C. Physicochemical studies of equine infectionus anemia virus. 3. Purification and electron microscopic observation of the virus.. Arch Gesamte Virusforsch 1969;28(3):348-60.
- Malmquist WA, Barnett D, Becvar CS. Production of equine infectious anemia antigen in a persistently infected cell line.. Arch Gesamte Virusforsch 1973;42(4):361-70.
- Ito Y, Kono Y, Kobayashi K. Electron microscopic observations of equine infectious anemia (EIA) virus in cultivated horse leukocytes. (Brief report).. Arch Gesamte Virusforsch 1969;28(3):411-6.
- Nakajima H, Yoshino T, Ushimi C. Equine infectious anemia virus from infected horse serum.. Infect Immun 1974 Sep;10(3):667-8.
- Kobayashi K, Kono Y. Propagation and titration of equine infectious anemia virus in horse leukocyte culture.. Natl Inst Anim Health Q (Tokyo) 1967 Spring;7(1):8-20.
- Nakajima H, Ushimi C. Immunodiffusion studies of purified equine infectious anemia virus.. Infect Immun 1971 Mar;3(3):373-7.
- McEwen CR. Tables for estimating sedimentation through linear concentration gradients of sucrose solution.. Anal Biochem 1967 Jul;20(1):114-49.
- LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent.. J Biol Chem 1951 Nov;193(1):265-75.
- Crawford TB, McGuire TC, Henson JB. Detection of equine infectious anemia virus in vitro by immunofluorescence.. Arch Gesamte Virusforsch 1971;34(4):332-9.
Citations
This article has been cited 7 times.- Sellon DC. Equine infectious anemia. Vet Clin North Am Equine Pract 1993 Aug;9(2):321-36.
- Sellon DC, Fuller FJ, McGuire TC. The immunopathogenesis of equine infectious anemia virus. Virus Res 1994 May;32(2):111-38.
- Cheevers WP, Ackley CM, Crawford TB. Structural proteins of equine infectious anemia virus. J Virol 1978 Dec;28(3):997-1001.
- Rice NR, Simek S, Ryder OA, Coggins L. Detection of proviral DNA in horse cells infected with equine infectious anemia virus. J Virol 1978 Jun;26(3):577-83.
- Weiland F, Matheka HD, Coggins L, Hatner D. Electron microscopic studies on equine infectious anemia virus (EIAV). Brief report. Arch Virol 1977;55(4):335-40.
- Cheevers WP, Archer BG, Crawford TB. Characterization of RNA from equine infectious anemia virus. J Virol 1977 Nov;24(2):489-97.
- Archer BG, Crawford TB, McGuire TC, Frazier ME. RNA-dependent DNA polymerase associated with equine infectious anemia virus. J Virol 1977 Apr;22(1):16-22.
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