Abstract
BackgroundCurrently, 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 FindingsHere, 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/SignificanceThe 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.
Highlights
Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus
We report that the enriched preparation of native Antigen 5 (Ag5) shows a very high and specific reactivity, both in western immunoblotting and in ELISA platforms, revealing promising perspectives for the development of sensitive immunodiagnostic tools
SDS-PAGE of sheep hydatid cyst fluid (HCF) A total of 4 different HCF samples collected from hydatid cysts, two localized in the lung and two in the liver of infected sheep, were concentrated and desalted by ultrafiltration and analyzed by SDS-PAGE under non-reducing and reducing conditions (Figure 1)
Summary
Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. The life cycle of this cestode involves dogs and other canids as the definitive hosts, while intermediate hosts are usually cattle, sheep, goats and pigs [1,2]. The greatest prevalence of CE in human and animal hosts has been recorded in the countries of the temperate zones, including parts of Eurasia, Australia, South America and Africa [3]. In most of these countries the disease has never been eradicated, exposing the population to the risk of reemergence in areas where CE was once believed to be controlled. Available immunodiagnostic tests lack standardization of the target antigen and, in turn, this is reflected on poor sensitivity and specificity of the serological diagnosis
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