Echinococcus granulosus protoscolex antigen used in serodiagnosis of hydatidosis by nano-gold dot-ELISA (original) (raw)
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Parasitology Research, 2010
Echinococcus granulosus causes human cystic echinococcosis as an important public health problem in many regions of the world. There are some problems in primary diagnosis such as cross-reaction with sera from patients with other parasitic disease in serological tests. The use of an appropriate source of antigenic material is a very important and crucial point in the improvement of the serodiagnostic features such as enzyme-linked immunosorbent assay (ELISA) method. We expressed and purified recombinant AgB of Echinococcus granulosus and used as antigen in ELISA method. Serum samples were given from 36 cystic hydatid disease patients that have been confirmed by surgical operation as well as 36 healthy individuals sera were tested by ELISA method using recombinant AgB and compared with commercial kit (Euroimmun) for specificity and sensitivities value. The sensitivity of 91.66% and specificity of 97.22% were determined by homemade kit.
Iranian journal of parasitology, 2010
Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test. DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individuals (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5) and tuberculosis (n=5) were examined using this recombinant antigen. Recombinant protein was purified by affinity chromatography using His-Tag column. After purification, recombinant protein was confirmed by western blot...
International Journal for Parasitology, 1993
BARBIER M., STERLA S., BA~TISTONI J. and NIETO A. 1993. High performance latex reagent for hydatid serology using an Echinococcus gramdosus lipoprotein antigen fraction purified from cyst fluid in one step. Internafional Journal for Parasitology 23: 565-572. A lipoprotein fraction from fertile bovine hydatid cyst fluid (FBHCF) was isolated by affinity chromatography on acrylic-heparin. The heparinbinding lipoprotein fraction (HBLF) was proved to be free from host immunoglobulins and albumin. lmmunoblotting with confirmed human hydatid sera showed that the major relevant diagnostic bands in FBHCF were also present in HBLF. HBLF or FBHCF were used to sensitize polystyrene latex particles. HBLF-latex showed both higher reactivity with positive hydatid human sera and higher batch-to-batch reproducibility than FBHCF-latex. Titration of sera from 119 surgically confirmed hydatid patients, 48 with other parasitic diseases, 6 with unrelated diseases and 37 healthy subjects was performed using HBLF-latex and conventional ELISA (using FBHCF). Sensitivity and specificity were 83 and 87% for ELISA and 87 and 88% for HBLF-latex.
Journal of Clinical Pathology, 1978
Preparations of the two most immunoreactive Echinococcus granulosus antigens (antigens 4 and 5) from sheep hydatid fluid, purified by a simplified method, and monospecific antisera against antigens 4 and 5, prepared by a new procedure, were used to measure the antigenic concentrations of antigens 4 and 5 in swine, sheep, and human hydatid fluids from pulmonary or hepatic cysts. Two bovine samples and two commercial preparations were also tested. The concentration of both antigens was significantly higher in sheep and human hydatid fluids than in swine hydatid fluid. The antigenic content of the two bovine samples and of the two commercial preparations was below the sensitivity level of the method employed. Independently of the species tested, the amount of Echinococcus antigens was greater in hepatic than in pulmonary cysts. The ratio between the concentrations of antigens 4 and 5 was constant at about 1:10 in the samples from various organs and from different species. When there were enough samples for statistical analysis a linear correlation was found between the contents of these two antigenic components but there was none between the amounts of proteins and the antigenic concentrations in the single cysts. Sheep hydatid fluid must therefore be considered the best source of antigenic material for diagnostic purposes even though in human cysts the antigenic fraction is less contaminated by serum proteins. We describe a reliable method of standardising antigenic material for the immunodiagnosis of hydatid disease.
Abstract Background: Cystic echinococcosis is an important zoonosis caused by the Echinococcus granulosus with a substantial impact in human and animal health in endemic areas. The purpose of the present study was serodiagnosis optimizing of dog echinococcosis in order to achieve a rapid diagnostic method. Methods: Eight dogs were challenged with protoscoleces in order to have positive echinococcosis serum and 2 two-month old puppies were used as uninfected controls. Colloidal gold was prepared by controlled reduction of a boiling solution of chloroauric acid (H [AuCl4]) with sodium citrate and labeled with recombinant EPC1. Dot immunogold filtration assay (DIGFA) was developed by coating rEPC1 labeled colloidal gold on nitrocellulose membrane. The canine sera, taken three times including, 15, 28 and 35 days post infection were tested. A total of 30 serum samples including 24 sera from 8 infected dogs and 6 sera from 2 puppies were comparatively detected with both DIGFA and ELISA. Results: Gold labeled antigen, showed a dark purple dot with agglutination particles in positive sera and light purple dot without agglutination in negative sera. Among 30 serum samples, 23 were positive and 7 were negative with DIGFA and 24 were positive, 6 were negative with ELISA. Conclusion: DIGFA as a rapid and simple procedure could be utilized in quickly diagnosis of echinococcosis.
Journal of Clinical Laboratory Analysis, 2001
Most serodiagnostic techniques have been evaluated for diagnosis of cystic hydatid disease caused by Echinococcus granulosus. Each, to varying degrees, has been shown to give false results, with considerable variation between laboratories. The comparative study was made concerning the sensitivity of the immunodiagnostic methods based on 58 sera from hydatid disease with different cyst locations. Latex agglutination, immunoelectrophoresis (IEP), and specific IgE, IgG enzyme-linked immunosorbent assay (ELISA) tests were studied. Specific IgG ELISA AgB (antigen B-rich fraction) was the most sensitive test (96.5%) and the least sensitive tests were specific IgE ELISA (24.1%) and IEP (25.8%).