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Evaluation of an ELISA for canine leishmaniasis immunodiagnostic using recombinant proteins.

Abstract
The present work describes the isolation and purification of two Leishmania chagasi (= syn. Leishmania infantum) recombinant proteins, rLci2B and rLci1A, and their use in the development of an immunoassay for the diagnostic of canine leishmaniasis. After protein expression and cell disruption, rLci2B was purified by immobilized metal affinity chromatography followed by size exclusion chromatography, whereas rLci1A, expressed as an inclusion body, was treated with urea and purified by anion-exchange chromatography. Homogeneities were ascertained by denaturing gel electrophoresis (MW (rLci2B) = 46,370; MW(rLci1A) = 88,400), isoelectric focusing (pI (rLci2B) = 5·91; pI (rLci1A) = 6·01) and Western blot. An indirect ELISA was developed using the purified antigens rLci2B and rLci1A and a leishmaniasis canine serum panel (n = 256). The ELISA showed 100% sensitivity and 95% specificity for rLci2B and 96% sensitivity and 92% specificity for rLci1A. The purified proteins did not present cross-reactivity with sera from dogs infected with Trypanosoma caninum, Babesia canis and Ehrlichia canis. Cross-reaction was verified with sera from dogs infected with Leishmania brasiliensis (11·7% for rLci2B and 2·9% for rLci1A). Based on ELISA results, it is suggested the use of rLci2B and rLci1A as antigens in an alternative serological assay for diagnostic of canine leishmania.
AuthorsC M de Souza, E D Silva, A P D Ano Bom, R C Bastos, H J Nascimento, J G da Silva Junior
JournalParasite immunology (Parasite Immunol) Vol. 34 Issue 1 Pg. 1-7 (Jan 2012) ISSN: 1365-3024 [Electronic] England
PMID21929686 (Publication Type: Evaluation Study, Journal Article)
Copyright© 2011 Blackwell Publishing Ltd.
Chemical References
  • Antibodies, Protozoan
  • Antigens, Protozoan
  • Recombinant Proteins
Topics
  • Animals
  • Antibodies, Protozoan (blood)
  • Antigens, Protozoan (genetics, isolation & purification)
  • Chromatography, Affinity (methods)
  • Chromatography, Ion Exchange (methods)
  • Cross Reactions
  • Dog Diseases (diagnosis)
  • Dogs
  • Enzyme-Linked Immunosorbent Assay (methods)
  • Leishmania infantum (genetics, immunology)
  • Leishmaniasis (diagnosis, veterinary)
  • Recombinant Proteins
  • Sensitivity and Specificity

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