To observed whether the specific IgG antibody test using ELISA was useful in diagnosis of presently ill patients of paragonimiasis, a total of 95 sera were tested. The sera were collected from 21 egg positive cases, 8 from positive reactors of intradermal test, 7 from Clonorchis infected, 9 from other parasitic diseases and 50 from apparently non-infected cases. By the result, the sensitivity of the test was 86% and the specificity was 100%. There were no cross reactions between Paragonimus antigen and other parastic infections. Specific IgG antibody test by micro-ELISA was concluded to be useful for mass screening of the presently ill paragonimiasis in the field.
parasitology-helminth-cestoda ; Spirometra sp. ; sparganum ; sparganosis ; immunology ; diagnosis ; ELISA

Seven cases of surgically proven sparganosis were serologically tested by means of micro ELISA for their specific IgG antibody levels. For that purpose, crude saline extract of spargana from snake, Natrix tigrina lateralis was prepared and used as antigen. The sparganosis sera were also tested with Paragonimus and Cysticercus antigens to observe the cross reactivity. A total of 71 sera from normal control, ectopic and pulmonary paragonimiasis, clonorchiasis, cysticercosis and Taenia saginata cases were also included. Except for one case of old calcified infection, all of 6 human sparganosis showed higher serum levels of specific IgG antibody when the differential point of positive reaction was set at the absorbance value of 0.25 (the sensitivity being 85.7%). In control and other helminthic infections, all except 3 cases of T. saginata infection showed negative reaction to sparganum antigen (the specificity being 95.7%). None of sparganosis cases showed cross reactivity to Paragonimus and Cysticercus antigens. Undiluted cerebrospinal fluid also showed high levels of antibody when central nervous system was invaded. The serologic diagnosis by means of micro-ELISA could be a useful tool in epidemiological study of human sparganosis in susceptible population, as well as in individual diagnosis.
parasitology-helminth-cestoda ; sparganosis ; sparganum ; Paragonimus ; Cysticercus ; ELISA ; immunology ; serology ; human

To observe the antigenic protein fractions in saline extract of Spirometra mansoni plerocercoid (sparganum), the crude extract was separated in reducing conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE). The proteins, transferred by electrophoresis to nitrocellulose paper, were reacted with sera from 15 surgically confirmed sparganosis and 24 cysticercosis patients for immunoblotting. Out of 30 identified protein bands in the extract, bands of 29 and 36 kilodaltons (kDa) were the strongest and the most frequently reacting with specific antibody (IgG) in sparganosis sera. Bands of higher molecular weight also reacted with the sera but their frequency of reactions was lower. Sera of cysticercosis reacted with different protein bands in saline extract of sparganum, but the cross reactions were observed in strong antigenic bands of 29 and 36 kDa.
parasitology-helminth-cestoda ; Spirometra mansoni ; plerocercoid ; sparganum ; antigen ; proteins ; sparganosis ; immunoblot ; immunology ; protein

We studied the serological reaction between various antigenic components from Cysticercus cellulosae and IgG antibodies in sera of cysticercosis, sparganosis, hydatidosis patients and normal humans by ELISA and EITB. In serological tests by ELISA, we recognized cross reaction of Cysticercus antigenic components with IgG antibodies in heterologous sera such as sparganosis and hydatidosis patients or normal humans. The crude antigenic components of Cysticercus showed lower ELISA sensitivity in homologous sera from cysticercosis patients than heterologous sera from hydatidosis patients. A total of 31 polypeptide bands with 260 KDa-22 KDa molecular weights were detected by SDS-PAGE, and 11 of them showed strong intensity. Total 22 components of them were recognized by IgG antibodies in cysticercosis patients sera. However, 12 of them were recognized also by normal human sera, 11 were by sparganosis sera, and 21 were by hydatidosis patients sera. The crude antigenic components of 104 KDa, 82 KDa, 72 KDa, 59 KDa and 34 KDa molecular weights were nonspecific ones, which cross-reacted with sera of either cysticercosis, sparganosis, hydatidosis patients or normal humans.
parasitology-helminth-cestoda ; Spirometra erinacei ; Taenia solium ; cysticercus ; sparganum ; immunology ; antigen

We purified specific 31/36 kDa antigenic molecules from sparganum in different intermediate hosts (snakes and mice) and analyzed their monosaccharide compositions. Compositional analysis showed that glucose and mannose concentrations were 2-3 fold higher in the 31/36 kDa molecule purified from snakes than those from mice. This result implies that antigenic glycoproteins of sparganum from snakes might be modified in mammalian sparganosis with respect to their carbohydrate composition.
Animals ; Antigens, Helminth/*chemistry ; Human ; Mice ; Monosaccharides/*analysis ; Snakes/parasitology ; Sparganum/immunology ; Spirometra/*immunology ; Support, Non-U.S. Gov't

After collecting calcareous corpuscles from plerocercoid of Spirometra mansoni (sparganum), we evaluated the antigenic values of calcareous corpuscles binding proteins obtained from the cyst fluid of Taenia solium metacestodes. Immunoblot analysis revealed that cysticercosis patient sera strongly recognized 10 and 95 kDa calcareous corpuscles binding proteins. This result demonstrated that calcareous corpuscles are bound with major secretory antigenic proteins, which is possibly involved in the secretory pathways of the 10 and 95 kDa proteins presenting in the cyst fluid of T. solium metacestodes.
Animals ; Antigens, Helminth/*analysis ; Carrier Proteins/*immunology ; Cysticercosis/diagnosis/immunology ; Helminth Proteins/*immunology ; Humans ; Immunoblotting/methods ; Molecular Weight ; Serologic Tests ; Sparganum ; Taenia solium/*chemistry/immunology

Competition ELISA test using sparganum specific monoclonal antibodies (Mab) was investigated to improve the diagnostic specificity of sparganosis. By cell fusion, one hybridoma clone secreting anti-sparganum specific Mab was selected (Sp-20), which reacted on bands of 32 kDa and 38 kDa. Sp-20 reacted on calcium corpuscles on IFA. By micro-ELISA, 16 of 17 sparganosis cases (95%) were found positive, but 1 of 18 clonorchiasis cases (5%), 4 of 16 cysticercosis cases (25%) and 2 of 16 normal controls (11%) showed false positive reactions. On the other hand, by competition ELISA using a sparganum specific Mab (Sp-20), 16 out of 17 (95%) of sparganosis cases were found positive, but 2 of 18 clonorchiasis cases (10%), 2 of 16 cysticercosis cases (12%), 3 of 16 paragonimiasis cases (18%) and 1 of 16 normal controls (6%) showed false positive reactions.
Animal ; Antibodies, Helminth ; Binding, Competitive ; Enzyme-Linked Immunosorbent Assay ; Human ; Sensitivity and Specificity ; Sparganosis/*diagnosis ; Sparganum/immunology ; Support, Non-U.S. Gov't

The author reported previously on separation of the outer tegument of the spargana (plerocercoids of Spirometra mansoni) using high concentration of urea solution. To determine which layer of the tegument is separated by this method, an electron microscopic analysis has been processed in this study. It was confirmed that the basement layer of the tegument is separated from the parenchyme of the sparganum. In addition, the antigenicity of the separated outer tegument against the human sparganosis patient sera was evaluated. Numerous antigenic proteins, including 16 and 55 kDa proteins, were noticed in the separated tegument; however, there were no diagnostic 31/36 kDa molecules in this tegument. The molecules reactive with the patient sera in the tegument are to be characterized in future studies.
Animal Structures/immunology/ultrastructure ; Animals ; Antigens, Helminth/chemistry/*immunology/isolation & purification ; Helminth Proteins/chemistry/*immunology/isolation & purification ; Humans ; Immunoblotting ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Molecular Weight ; Sparganum/*immunology/*ultrastructure

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.
Animals ; Antigens, Helminth/*analysis/chemistry/immunology/metabolism ; Carbohydrates/analysis/immunology ; Chromatography, Affinity ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Epitopes/analysis/immunology ; Glucosaminidase/metabolism ; Human ; Peptide-N4- (N-acetyl-beta-glucosaminyl) Asparagine Amidase/metabolism ; Periodic Acid/chemistry ; Sparganosis/*parasitology ; Sparganum/immunology/metabolism ; Spirometra/immunology/*metabolism ; Support, Non-U.S. Gov't

A 29-year-old Korean woman visited the Department of Surgery in MizMedi Hospital with a palpable itching mass on the right breast that had existed for the past 7 months. She had no history to eat either frogs or snakes, but had the history of drinking impure water. Sonography revealed a serpiginous hypoechoic tubular structure associated with partial fat necrosis in breast parenchymal layer and subcutaneous fat layer. It also revealed oval cystic lesions. At operation, an ivory white opaque ribbon-like worm that measured 16.5 cm in length and 0.5 cm in width was extracted. Anti-sparganum specific serum IgG level in the patient's serum (absorbance = 0.71), measured by ELISA, was found to be significantly higher than those of normal controls (cut off point = 0.21). Sonography and ELISA appear to be helpful to diagnose sparganosis. Breast sparganosis is rarely found throughout the world.
Adult ; Animals ; Antibodies, Helminth/blood ; Breast/*parasitology ; Breast Diseases/diagnosis/*parasitology ; Enzyme-Linked Immunosorbent Assay ; Female ; Human ; Immunoglobulin G/blood ; Sparganosis/diagnosis/*parasitology ; Sparganum/immunology/isolation & purification ; Ultrasonography, Mammary