An Easy and Efficient Method for Native and Immunoreactive Echinococcus granulosus Antigen 5 Enrichment from Hydatid Cyst Fluid
Caricamento...
Data
2014-08-13
Autori
Pagnozzi, Daniela
Biosa, Grazia
Addis, Maria Filippa
Mastrandrea, Scilla
Masala, Giovanna
Uzzau, Sergio
Titolo del periodico
ISSN
Titolo del volume
Editore
Public Library of Science
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.
Methodology/Principal Findings: 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/Significance: 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.
Descrizione
Keywords
antigens , diagnostic medicine , echinococcosis , enzyme-linked immunoassays , serum proteins , membrane proteins , sheep , size-exclusion chromatography
Citazione
Pagnozzi D, Biosa G, Addis MF, Mastrandrea S, Masala G, et al. (2014) An Easy and Efficient Method for Native and Immunoreactive Echinococcus granulosus Antigen 5 Enrichment from Hydatid Cyst Fluid. PLoS ONE 9(8): e104962. doi:10.1371/journal.pone.0104962