title: Analysis of an expressed sequence tag library from Dicrocoelium dentriticum.
abstract: Dicrocoeliosis caused by Dicrocoelium dendriticum is an important liver disease, which affects ruminants all around the world. Despite the significant economic losses caused by this trematode, molecular knowledge is very scarce. In fact, there is no information in the expressed sequence tag (EST) database about the parasite. Furthermore, the immunological diagnosis of dicrocoeliosis remains unsatisfactory, and there aren't available recombinant proteins that could be tested in the diagnosis. For this reason a cDNA library was constructed with mRNA extracted from D. dendriticum adults for first time. A random preliminary screening of 230 phage plaques from the library resulted in the identification of 173 new EST. The deduced proteins expressed by these genes have been described as possible vaccine targets in other trematodes, and/or as relevant diagnosis antigens. Then, our goal was to identify D. dentriticum diagnosis genes to be used as recombinant antigens in the specific immunological diagnosis of the trematodoses. A D. dendriticum cDNA encoding an 8-kDa recombinant protein has been cloned, expressed in Escherichia coli and evaluated in dicrocoeliosis diagnosis using both Western Blot and enzyme-linked immunosorbent assay (ELISA). The recombinant expression molecule has demonstrated its value as a diagnosis antigen of dicrocoeliosis, able to discriminate between positive and controls on day 30 post infection. This is the first research conducted for identification and characterization of D. dendriticum ESTs, which can serve as a starting point for future research on immunodiagnosis and immunoprofilaxis of dicrocoeliosis.
authors.affiliation: Departamento de Sanidad Animal, Instituto de Ganadería de Montaña, Consejo Superior de Investigaciones Científicas (CSIC-ULE), 24346 Grulleros, León, Spain.
authors.forename: A M
authors.initials: AM
authors.lastname: Martínez-Ibeas
authors.forename: M J
authors.initials: MJ
authors.lastname: Perteguer
authors.forename: C
authors.initials: C
authors.lastname: González-Lanza
authors.forename: T
authors.initials: T
authors.lastname: Gárate
authors.forename: M Y
authors.initials: MY
authors.lastname: Manga-González
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D019295:Computational Biology; D003429:Cross Reactions; D018076:DNA, Complementary / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry; D004011:Dicrocoeliasis / Q000175:diagnosis* / Q000469:parasitology; D004013:Dicrocoelium / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D020224:Expressed Sequence Tags; D015870:Gene Expression; D015723:Gene Library; D007106:Immune Sera / Q000276:immunology; D008099:Liver / Q000469:parasitology; D008297:Male; D008969:Molecular Sequence Data; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000175:diagnosis / Q000469:parasitology
chemical_list: D000947:Antigens, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D007106:Immune Sera; D018510:RNA, Helminth; D011994:Recombinant Proteins
keywords: 8-kDa protein; Diagnosis; Dicrocoelium dendriticum; Expressed sequence tag (EST); cDNA library