Serum antibodies from a subset of horses positive for Babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection

Peter O. Awinda, Robert H. Mealey, Laura B A Williams, Patricia A Conrad, Andrea E. Packham, Kathryn E. Reif, Juanita F. Grause, Angela M. Pelzel-McCluskey, Chungwon Chung, Reginaldo G. Bastos, Lowell S. Kappmeyer, Daniel K. Howe, SallyAnne L. Ness, Donald P. Knowles, Massaro W. Ueti

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Tick-borne pathogens that cause persistent infection are of major concern to the livestock industry because of transmission risk from persistently infected animals and the potential economic losses they pose. The recent reemergence of Theileria equi in the United States prompted a widespread national survey resulting in identification of limited distribution of equine piroplasmosis (EP) in the U.S. horse population. This program identified Babesia caballi-seropositive horses using rhoptry-associated protein 1 (RAP-1)-competitive enzyme-linked immunosorbent assay (cELISA), despite B. caballi being considered nonendemic on the U.S. mainland. The purpose of the present study was to evaluate the suitability of RAP-1-cELISA as a single serological test to determine the infection status of B. caballi in U.S. horses. Immunoblotting indicated that sera from U.S. horses reacted with B. caballi lysate and purified B. caballi RAP-1 protein. Antibody reactivity to B. caballi lysate was exclusively directed against a single ∼50-kDa band corresponding to a native B. caballi RAP-1 protein. In contrast, sera from experimentally and naturally infected horses from regions where B. caballi is endemic bound multiple proteins ranging from 30 to 50 kDa. Dilutions of sera from U.S. horses positive by cELISA revealed low levels of antibodies, while sera from horses experimentally infected with B. caballi and from areas where B. caballi is endemic had comparatively high antibody levels. Finally, blood transfer from seropositive U.S. horses into naive horses demonstrated no evidence of B. caballi transmission, confirming that antibody reactivity in cELISA-positive U.S. horses was not consistent with infection. Therefore, we conclude that a combination of cELISA and immunoblotting is required for the accurate serodiagnosis of B. caballi.

Original languageEnglish (US)
Pages (from-to)1752-1757
Number of pages6
JournalClinical and Vaccine Immunology
Volume20
Issue number11
DOIs
StatePublished - Nov 2013

Fingerprint

Babesia
Immunosorbents
Horses
Pattern recognition
Assays
Enzyme-Linked Immunosorbent Assay
Antibodies
Enzymes
Infection
Serum
Proteins
Serologic Tests
Immunoblotting
Pathogens
Farms
Theileria
Dilution
Babesiosis
Animals
Blood

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Immunology
  • Immunology and Allergy
  • Microbiology (medical)

Cite this

Serum antibodies from a subset of horses positive for Babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection. / Awinda, Peter O.; Mealey, Robert H.; Williams, Laura B A; Conrad, Patricia A; Packham, Andrea E.; Reif, Kathryn E.; Grause, Juanita F.; Pelzel-McCluskey, Angela M.; Chung, Chungwon; Bastos, Reginaldo G.; Kappmeyer, Lowell S.; Howe, Daniel K.; Ness, SallyAnne L.; Knowles, Donald P.; Ueti, Massaro W.

In: Clinical and Vaccine Immunology, Vol. 20, No. 11, 11.2013, p. 1752-1757.

Research output: Contribution to journalArticle

Awinda, PO, Mealey, RH, Williams, LBA, Conrad, PA, Packham, AE, Reif, KE, Grause, JF, Pelzel-McCluskey, AM, Chung, C, Bastos, RG, Kappmeyer, LS, Howe, DK, Ness, SL, Knowles, DP & Ueti, MW 2013, 'Serum antibodies from a subset of horses positive for Babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection', Clinical and Vaccine Immunology, vol. 20, no. 11, pp. 1752-1757. https://doi.org/10.1128/CVI.00479-13
Awinda, Peter O. ; Mealey, Robert H. ; Williams, Laura B A ; Conrad, Patricia A ; Packham, Andrea E. ; Reif, Kathryn E. ; Grause, Juanita F. ; Pelzel-McCluskey, Angela M. ; Chung, Chungwon ; Bastos, Reginaldo G. ; Kappmeyer, Lowell S. ; Howe, Daniel K. ; Ness, SallyAnne L. ; Knowles, Donald P. ; Ueti, Massaro W. / Serum antibodies from a subset of horses positive for Babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection. In: Clinical and Vaccine Immunology. 2013 ; Vol. 20, No. 11. pp. 1752-1757.
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abstract = "Tick-borne pathogens that cause persistent infection are of major concern to the livestock industry because of transmission risk from persistently infected animals and the potential economic losses they pose. The recent reemergence of Theileria equi in the United States prompted a widespread national survey resulting in identification of limited distribution of equine piroplasmosis (EP) in the U.S. horse population. This program identified Babesia caballi-seropositive horses using rhoptry-associated protein 1 (RAP-1)-competitive enzyme-linked immunosorbent assay (cELISA), despite B. caballi being considered nonendemic on the U.S. mainland. The purpose of the present study was to evaluate the suitability of RAP-1-cELISA as a single serological test to determine the infection status of B. caballi in U.S. horses. Immunoblotting indicated that sera from U.S. horses reacted with B. caballi lysate and purified B. caballi RAP-1 protein. Antibody reactivity to B. caballi lysate was exclusively directed against a single ∼50-kDa band corresponding to a native B. caballi RAP-1 protein. In contrast, sera from experimentally and naturally infected horses from regions where B. caballi is endemic bound multiple proteins ranging from 30 to 50 kDa. Dilutions of sera from U.S. horses positive by cELISA revealed low levels of antibodies, while sera from horses experimentally infected with B. caballi and from areas where B. caballi is endemic had comparatively high antibody levels. Finally, blood transfer from seropositive U.S. horses into naive horses demonstrated no evidence of B. caballi transmission, confirming that antibody reactivity in cELISA-positive U.S. horses was not consistent with infection. Therefore, we conclude that a combination of cELISA and immunoblotting is required for the accurate serodiagnosis of B. caballi.",
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