Quantitation of Leishmania infantum in tissues of infected BALB/c mouse by sandwich ELISA
Autor: | Bernard Ferrua, C. Roptin, Y Le Fichoux, J. Kubar, D Rousseau, I Suffia |
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Rok vydání: | 2001 |
Předmět: |
Mice
Inbred BALB C biology BALB/c Mouse Clinical Biochemistry Immunology Spleen Enzyme-Linked Immunosorbent Assay biology.organism_classification Molecular biology Sensitivity and Specificity Medical Laboratory Technology Microtiter plate Mice medicine.anatomical_structure Antigen biology.protein medicine Immunology and Allergy Parasite hosting Animals Leishmania infantum Amastigote Peroxidase |
Zdroj: | Journal of immunoassayimmunochemistry. 22(2) |
ISSN: | 1532-1819 |
Popis: | In this report, a sandwhich ELISA was developed to quantify spleen and liver burdens from L. infantum-infected BALB/c mice. Amastigote antigens obtained following Nonidet P40 extraction of parasite-harbouring tissues were captured by anti-L. infantum human IgG insolubilized onto microtiter plate and subsequently revealed with anti-L. infantum F(ab)' fragments labelled with peroxidase. The method was easy to perform, precise and capable to specifically and accurately detect 5 x 10(4) amastigotes/100 mg tissue. Parasite burdens from infected BALB/c mice, in various conditions, were measured by ELISA and Giemsa-stained touch imprint reference methods, and compared. Both techniques agreed well with close values for liver burdens, but the spleen loads measured by the ELISA were, on average, 10.7 times higher than those calculated from imprints. This difference was attributed partly to the underestimation brought by Stauber's formula. However, it did not preclude the usefulness of this newly developed test, since results obtained in kinetics studies and evaluation of the efficiency of leishmanicidal drugs allowed us to draw identical conclusions. |
Databáze: | OpenAIRE |
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