title: An easy and efficient method for native and immunoreactive Echinococcus granulosus antigen 5 enrichment from hydatid cyst fluid.
abstract: BACKGROUND
  Currently, the serodiagnosis of cystic echinococcosis relies mostly on crude Echinococcus granulosus hydatid cyst fluid as the antigen. Consequently, available immunodiagnostic tests lack standardization of the target antigen and, in turn, this is reflected on poor sensitivity and specificity of the serological diagnosis.
  
  
  RESULTS
  Here, a chromatographic method enabling the generation of highly enriched Antigen 5 (Ag5) is described. The procedure is very easy, efficient and reproducible, since different hydatid cyst fluid (HCF) sources produced very similar chromatograms, notwithstanding the clearly evident and extreme heterogeneity of the starting material. In addition, the performance of the antigen preparation in immunological assays was preliminarily assessed by western immunoblotting and ELISA on a limited panel of cystic echinococcosis patients and healthy controls. Following western immunoblotting and ELISA experiments, a high reactivity of patient sera was seen, with unambiguous and highly specific results.
  
  
  CONCLUSIONS
  The methods and results reported open interesting perspectives for the development of sensitive diagnostic tools to enable the timely and unambiguous detection of cystic echinococcosis antibodies in patient sera.
authors.affiliation: Porto Conte Ricerche Srl, Tramariglio, Alghero, Sassari, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Pagnozzi
authors.affiliation: Porto Conte Ricerche Srl, Tramariglio, Alghero, Sassari, Italy.
authors.forename: Grazia
authors.initials: G
authors.lastname: Biosa
authors.affiliation: Porto Conte Ricerche Srl, Tramariglio, Alghero, Sassari, Italy.
authors.forename: Maria Filippa
authors.initials: MF
authors.lastname: Addis
authors.affiliation: Centro Nazionale di Riferimento per l'Echinococcosi, IZS "G. Pegreffi", Sassari, Italy; Unità Operativa Complessa di Malattie Infettive, Azienda Ospedaliera Universitaria, Sassari, Italy.
authors.forename: Scilla
authors.initials: S
authors.lastname: Mastrandrea
authors.affiliation: Centro Nazionale di Riferimento per l'Echinococcosi, IZS "G. Pegreffi", Sassari, Italy.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Masala
authors.affiliation: Porto Conte Ricerche Srl, Tramariglio, Alghero, Sassari, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Uzzau
mesh_terms: D000818:Animals; D004443:Echinococcosis / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary; D048209:Echinococcus granulosus / Q000276:immunology / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D053719:Tandem Mass Spectrometry
chemical_list: D006023:Glycoproteins; C514021:antigen 5, Echinococcus granulosus