Gastroenterology and hepatology from bed to bench.
Stepwise purification, characterization, and diagnostic evaluation of antigen B isolated from Echinococcus granulosus hydatid cyst fluid for indirect ELISA detection of human cystic echinococcosis: Echinococcus granulosus hydatid cyst fluid for indirect ELISA detection of human cystic echinococcosis
By Mona Maleki, Sara Nemati, Farzaneh Hosseini and 2 others.
Abstract
Background
Cystic echinococcosis (CE) is a globally significant zoonosis requiring improved, cost-effective serodiagnosis. Accurate detection is often hindered by cross-reactive host contaminants in crude hydatid cyst fluid (HCF). This study aimed to purify and evaluate the immunogenicity of native Antigen B (AgB) extracted from sheep HCF for indirect ELISA detection of human CE.
Methods
Native AgB was purified from sheep HCF using sequential dialysis, ultracentrifugation, ammonium sulfate precipitation, and Amicon ultrafiltration. Protein concentration was monitored throughout purification. Purity was assessed by electrophoresis and gel permeation chromatography (GPC). Immunoreactivity was evaluated by indirect ELISA using sera from CE patients and healthy controls previously validated with a commercial kit. Serum dilution optimization and assay cut-off values were established using serial dilution experiments.