Immunodiagnosis of fasciolosis using recombinant procathepsin L cystein proteinase.
Cathepsin L1, a cysteine protease secreted by the gastrodermis of juvenile and adult Fasciola hepatica, was expressed in Escherichia coli as a fusion protein containing the proregion, supplied with six histidyl residues at the N-terminal end (rproCL1). In this study we tested its potential as antigen for the serologic diagnosis of F. hepatica infections by enzyme-linked immunosorbent assay (ELISA). The analyzed human sera included 16 positive samples, 99 negative controls and 111 from individuals affected by other parasitic and non parasitic diseases. The sensitivity and specificity of the rproCL1-ELISA were 100%. We also assessed the ability to detect antibodies in sera from 10 experimentally infected sheep, obtaining preliminary results that shown a response since the third week post infection in all the studied animals. Therefore, the recombinant rproCL1-based ELISA could be a standardized test for the accurate diagnosis of fasciolosis.
Carnevale
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Departamento de Parasitología, ANLIS Dr. Carlos G. Malbrán, Av. Vélez Sársfield 563, (1281), Buenos Aires, Argentina. jorsil@overnet.com.ar
Rodríguez
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Guarnera
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EA
Carmona
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Tanos
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Angel
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D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000097:blood*; D056668:Cathepsin L; D002403:Cathepsins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005210:Fasciola hepatica / Q000276:immunology* / Q000302:isolation & purification; D005211:Fascioliasis / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D018510:RNA, Helminth; D011993:Recombinant Fusion Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology
D000909:Antibodies, Helminth; D004792:Enzyme Precursors; D007074:Immunoglobulin G; D018510:RNA, Helminth; D011993:Recombinant Fusion Proteins; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C063905:procathepsin L; D056668:Cathepsin L