Serodiagnostic potential of chemically synthesized glycosphingolipid antigens in an enzyme-linked immunosorbent assay for alveolar echinococcosis

2006 ◽  
Vol 80 (4) ◽  
pp. 387-391 ◽  
Author(s):  
K. Yamano ◽  
N. Hada ◽  
T. Yamamura ◽  
T. Takeda ◽  
H. Honma ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Immunosorbent Assay ◽  
Echinococcus Multilocularis ◽  
Cystic Echinococcosis ◽  
Alveolar Echinococcosis ◽  
Carbohydrate Antigen ◽  
Single Component ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Specific Reactions

AbstractIn the serodiagnosis of alveolar echinococcosis, the detection of specific reactions against not only protein but also carbohydrate antigen is useful and both antigens supplement each other. Though recombinant protein antigens have recently advanced, the preparation of carbohydrate antigen still depends on extraction from crude antigens. In the latter case, it is not conventional to obtain carbohydrate antigen as a single component for examination and research. Therefore, chemically synthesized carbohydrate antigens were prepared for serodiagnosis by the enzyme-linked immunosorbent assay (ELISA). Four antigens with the structure of glycosphingolipids fromEchinococcus multiloculariswere examined and one antigen, Galβ1-6(Fucα1-3)Galβ1-6Galβ1-ceramide, was found to show significant serodiagnostic potential in differentiating alveolar from cystic echinococcosis.

scholarly journals Synthesis of the Carbohydrate Moiety of Glycoproteins from the Parasite Echinococcus granulosus and Their Antigenicity against Human Sera

Molecules ◽  
2021 ◽  
Vol 26 (18) ◽  
pp. 5652
Author(s):  
Noriyasu Hada ◽  
Tokio Morita ◽  
Takashi Ueda ◽  
Kazuki Masuda ◽  
Hiromi Nakane ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Immunosorbent Assay ◽  
Cystic Echinococcosis ◽  
Alveolar Echinococcosis ◽  
Total Yield ◽  
Carbohydrate Moiety ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Sera

Stereocontrolled syntheses of biotin-labeled oligosaccharide portions containing the carbohydrate moiety of glycoprotein from Echinococcus granulosus have been accomplished. Trisaccharide Galβ1-3Galβ1-3GalNAcα1-R (A), tetrasaccharide Galα1-4Galβ1-3Galβ1-3GalNAcα1-R (B), and pentasaccharide Galα1-4Galβ1-3Galβ1-3Galβ1-3GalNAcα1-R (C), (R = biotinylated probe) were synthesized by stepwise condensation and/or block synthesis by the use of 5-(methoxycarbonyl)pentyl 2-azido-4,6-O-benzylidene-2-deoxy-α-d-galactopyranoside as a common glycosyl acceptor. The synthesis of the tetrasaccharide and the pentasaccharide was improved from the viewpoint of reducing the number of synthetic steps and increasing the total yield by changing from stepwise condensation to block synthesis. Moreover, hexasaccharide E, which contains the oligosaccharide sequence which occurs in E. granulosus, was synthesized from trisaccharide D. We examined the antigenicity of these five oligosaccharides by an enzyme-linked immunosorbent assay (ELISA). Although compounds of C–E did not exhibit antigenicity against cystic echinococcosis (CE) patient sera, compounds B, D, and E showed good serodiagnostic potential for alveolar echinococcosis (AE).

scholarly journals Detection of Echinococcus Multilocularis Infection in a Colony of Cynomolgus Monkeys (Macaca Fascicularis) using Serology and Ultrasonography

2005 ◽  
Vol 17 (2) ◽  
pp. 183-186 ◽  
Author(s):  
Patrick Rehmann ◽  
Andrea Gröne ◽  
Bruno Gottstein ◽  
Jörg Völlm ◽  
Heinz Sager ◽  
...  
Keyword(s):  
Bile Duct ◽  
Immunosorbent Assay ◽  
Echinococcus Multilocularis ◽  
Macaca Fascicularis ◽  
Alveolar Echinococcosis ◽  
Postmortem Examination ◽  
The Other ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cynomolgus Monkeys ◽  
Serological Examination

Five animals in a colony of cynomolgus monkeys ( Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis.

Diagnosis of alveolar echinococcosis using immunoblotting with plural low molecular weight antigens

2009 ◽  
Vol 83 (1) ◽  
pp. 57-61 ◽  
Author(s):  
K. Yamano ◽  
A. Goto ◽  
M. Miyoshi ◽  
K. Furuya ◽  
Y. Sawada ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Alveolar Echinococcosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Screening ◽  
Screening Process ◽  
Medical Practitioners ◽  
Specific Reactions ◽  
Hospital Outpatients ◽  
Higher Sensitivity

AbstractAlveolar echinococcosis (AE) is endemic to Hokkaido, Japan. For the past 20 years, detection of AE among inhabitants has involved serological screening using an enzyme-linked immunosorbent assay (ELISA) followed by Western blotting (WB). Between the years 1987 and 2000, antigens targeted on 66, 55 and 30–35 kDa bands were routinely used in the WB step of AE diagnosis. However, since 2001 diagnosis has been dependent on three smaller molecular weight antigens (26–28, 18 and 7–8 kDa). Due to its higher sensitivity, this improved WB approach has been used as a confirmation step in the screening process and also for the testing of suspected AE cases in hospital outpatients. Using the improved WB technique, a total of 1745 serum samples were examined in 2001–2006 with 81 patients detected and registered with AE. Interestingly, sera from 76 of the 81 diagnosed AE patients (93.8%) demonstrated reactivity with all three antigens. However, sera from the remaining five patients (6.2%) demonstrated no reactivity with the 18 kDa antigen, even though they exhibited clearly detectable levels of reactivity with the 26–28 and 7–8 kDa bands. These results suggest that medical practitioners need to pay particular attention to the specific reactions to some different diagnostic antigens to minimize the risk of misdiagnosing AE patients. In turn, these results may also provide important diagnostic information for cystic echinococcosis (CE).

scholarly journals Alveolar Echinococcosis in Children

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Emilija Jonaitytė ◽  
Martynas Judickas ◽  
Eglė Tamulevičienė ◽  
Milda Šeškutė
Keyword(s):  
Surgical Treatment ◽  
Current Literature ◽  
Echinococcus Multilocularis ◽  
Cystic Echinococcosis ◽  
Alveolar Echinococcosis ◽  
Pediatric Patients ◽  
Treatment Options ◽  
Clinical Manifestations ◽  
Diagnostic Methods

Alveolar echinococcosis (AE) is an infectious zoonotic disease that is caused by Echinococcus multilocularis. The disease is generally identified accidentally because of the long asymptomatic period, has a malignant behaviour, and mainly occurs in the liver. Usually it is diagnosed in adults and is very rare in pediatric patients. We report two cases of AE and 1 differential case between AE and cystic echinococcosis (CE) in children: two of them had lesions in the liver and one had rare extrahepatic presentation of a cyst in the spleen. All our patients received chemotherapy with albendazole because surgical treatment was not recommended. The children were followed-up from 10 to 30 months and no significant improvement was seen. In this report we discuss the difficulties we faced in the treatment and follow-up of these patients. We also review the main clinical manifestations, general diagnostic methods, and treatment options of AE according to the current literature.

Immuno-enzymatic assay for the detection of Schistosoma mansoni antigens in serum of mice harbouring bisexual or unisexual light worm infections

1979 ◽  
Vol 53 (3) ◽  
pp. 189-194 ◽  
Author(s):  
A. W. Ferreira ◽  
A. L. M. Caldini ◽  
S. Hoshino-Shimizu ◽  
M. E. Camargo
Keyword(s):  
Schistosoma Mansoni ◽  
Immunosorbent Assay ◽  
Enzymatic Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Male Worm ◽  
Antibody Technique ◽  
Post Exposure ◽  
Double Antibody

ABSTRACTEnzyme-linked immunosorbent assay (ELISA) double antibody technique was standardized for detecting S. mansoni Polysaccharide and protein antigens in serum of infected mice. Anti-sera specific for either worm components were obtained in sheep and peroxidase conjugates prepared from each serum. The immunoenzimatic test for protein could detect as little as 6μg/ml antigens, and the test for polysaccharides 3 μg/ml.Both bisexual and unisexual male worm low infections were produced, and studied for as long as 27 weeks post-exposure. Worm components were found in serum from both types of infections and in progressively higher percentages of animals until the end of the 27 weeks observations period. For unisexual male infections this percentage reached from about 50% to 60% of mice, and 100% for bisexual infections. Significantly higher antigen concentrations in serum were found at 27 weeks for bisexual infections, no antigen increase being detected in relation to starting egg secretion, which occurred at 5 week infections.

scholarly journals Molecular Expression of a Cysteine Proteinase of Clonorchis sinensis and Its Application to an Enzyme-Linked Immunosorbent Assay for Immunodiagnosis of Clonorchiasis

2004 ◽  
Vol 11 (2) ◽  
pp. 411-416 ◽  
Author(s):  
Isao Nagano ◽  
Fuquan Pei ◽  
Zhiliang Wu ◽  
Jun Wu ◽  
Huier Cui ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Adult Worm ◽  
Immunosorbent Assay ◽  
Fasciola Hepatica ◽  
Cysteine Proteinase ◽  
Clonorchis Sinensis ◽  
Recombinant Antigen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Predicted Amino Acid Sequence ◽  
Crude Extracts

ABSTRACT We produced a recombinant cysteine proteinase of Clonorchis sinensis and tested its value as an antigen for serologic diagnosis of C. sinensis infections. The predicted amino acid sequence of the cysteine proteinase of C. sinensis was 58, 48, and 40% identical to those of cathepsin L cysteine proteinases from Paragonimus westermani, Schistosoma japonicum, and Fasciola hepatica, respectively. Western blotting analysis showed that sera from patients infected with C. sinensis strongly reacted with the recombinant protein and that sera from patients infected with S. japonicum weakly reacted with the recombinant protein. Antibody against the recombinant protein stained proteins migrating at about 37 and 28 kDa in C. sinensis adult worm crude extracts. Immunostaining revealed that the cysteine proteinase of C. sinensis was located in the intestinal epithelial cells of the adult parasite and in intrauterine eggs. The specificity and sensitivity of the recombinant antigen or C. sinensis adult worm crude extracts were assessed by an enzyme-linked immunosorbent assay (ELISA) using serum samples from humans infected with different parasites, including 50 patients with clonorchiasis, and negative controls. The sensitivities of the ELISA with the recombinant antigen and C. sinensis adult worm crude extracts were 96 and 88%, respectively. The specificities of the ELISA with the recombinant antigen and C. sinensis adult worm crude extracts were 96.2 and 100%, respectively. The results suggested that the recombinant cysteine proteinase-based ELISA could provide a highly sensitive and specific assay for diagnosis of clonorchiasis.

scholarly journals Detection of Antibodies against Epidermodysplasia Verruciformis-Associated Canine Papillomavirus 3 in Sera of Dogs from Europe and Africa by Enzyme-Linked Immunosorbent Assay

2008 ◽  
Vol 16 (1) ◽  
pp. 66-72 ◽  
Author(s):  
C. E. Lange ◽  
K. Tobler ◽  
C. Favrot ◽  
M. Müller ◽  
J. O. Nöthling ◽  
...  
Keyword(s):  
South Africa ◽  
Fusion Protein ◽  
Immunosorbent Assay ◽  
Clinical Manifestations ◽  
Epidermodysplasia Verruciformis ◽  
Capsid Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Specific Antibodies

ABSTRACT The role of papillomaviruses (PVs) in the development of canine cancers is controversial. However, recently a novel canine PV (CPV3) was detected in a dog affected with a condition reminiscent of epidermodysplasia verruciformis (EV). The aim of the present study was to investigate the seroprevalence of CPV3 by using generic enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against either canine oral PV (COPV) or CPV3. Therefore, the capsid proteins of both PV types were expressed as glutathione S-transferase fusion protein antigens and adsorbed to glutathione-casein-coated ELISA plates. After showing that PV type-specific antibodies could be detected in the sera from dogs with confirmed COPV or CPV3 infection, CPV3- and COPV-seropositive samples were detected in two sets of canine sera collected in Switzerland and South Africa, respectively. We found specific antibodies against COPV and CPV3 among the tested sera and also a large number that were positive for both antigens. The seroprevalences of PV antibodies of 21.9% (COPV) and 26.9% (CPV3) among the tested dogs from South Africa were higher than those among the dogs from Switzerland at 10.5% (COPV) and 1.3% (CPV3). Our data suggest a need for further CPV-related seroepidemiological surveys in different countries, especially in the context of clinical manifestations and possible breed predispositions. For this purpose, the newly developed ELISAs can be a useful tool.

scholarly journals Heterogeneous Antibody Responses in Tuberculosis

1998 ◽  
Vol 66 (8) ◽  
pp. 3936-3940 ◽  
Author(s):  
Konstantin Lyashchenko ◽  
Roberto Colangeli ◽  
Michel Houde ◽  
Hamdan Al Jahdali ◽  
Dick Menzies ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Antibody Response ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Antigen Recognition ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Tuberculosis Patients ◽  
Specific Antibodies

ABSTRACT Antibody responses during tuberculosis were analyzed by an enzyme-linked immunosorbent assay with a panel of 10 protein antigens of Mycobacterium tuberculosis. It was shown that serum immunoglobulin G antibodies were produced against a variety of M. tuberculosis antigens and that the vast majority of sera from tuberculosis patients contained antibodies against one or more M. tuberculosis antigens. The number and the species of serologically reactive antigens varied greatly from individual to individual. In a given serum, the level of specific antibodies also varied with the antigen irrespective of the total number of antigens recognized by that particular serum. These findings indicate that person-to-person heterogeneity of antigen recognition, rather than recognition of particular antigens, is a key attribute of the antibody response in tuberculosis.

scholarly journals Cloning and Expression of an Immunogenic Membrane-Associated Protein of Helicobacter hepaticus for Use in an Enzyme-Linked Immunosorbent Assay

1999 ◽  
Vol 6 (5) ◽  
pp. 745-750 ◽  
Author(s):  
Robert S. Livingston ◽  
Lela K. Riley ◽  
Reuel R. Hook ◽  
Cynthia L. Besch-Williford ◽  
Craig L. Franklin
Keyword(s):  
Recombinant Protein ◽  
Fusion Protein ◽  
Immunosorbent Assay ◽  
Outer Membrane Proteins ◽  
Cloning And Expression ◽  
Signal Peptidase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Predicted Amino Acid Sequence ◽  
Helicobacter Hepaticus ◽  
Western Blots

ABSTRACT Helicobacter hepaticus is a bacterial pathogen that causes chronic active hepatitis and inflammatory bowel disease in mice. The purpose of this study was to develop a recombinant antigen-based enzyme-linked immunosorbent assay (ELISA) to detect H. hepaticus-infected mice. A genomic library of H. hepaticus was constructed and was screened with sera fromH. hepaticus-infected mice. A 459-bp open reading frame that coded for an 18-kDa immunoreactive protein, MAP18, was identified. The gene had high identity with genes coding for outer membrane proteins of other bacteria, and the predicted amino acid sequence of MAP18 had a putative membrane-trafficking signal sequence and a putative signal peptidase II cleavage site. The recombinant protein was expressed in Escherichia coli as a glutathioneS-transferase (GST) fusion protein, GST-MAP18, and purified by affinity chromatography. The 44-kDa fusion protein was detected on Western blots probed with sera from H. hepaticus-infected mice but was not detected on blots probed with sera from mice infected with Helicobacter muridarum or Helicobacter bilis or with sera from mice free of Helicobacterinfection. The GST-MAP18 fusion protein was used as an antigen in an ELISA to detect anti-H. hepaticus antibodies in sera from infected mice. This ELISA was compared to an H. hepaticus-specific ELISA that uses a detergent extract ofH. hepaticus as the antigen. Sera from mice naturally and experimentally infected with H. hepaticus, H. bilis, or H. muridarum and sera from mice free ofHelicobacter infection were evaluated. Both ELISAs performed with a high specificity (98%); however, the detergent extract-based ELISA performed with a higher sensitivity (89%) than the recombinant protein-based ELISA (sensitivity, 66%). These data indicate that H. hepaticus carries a gene that encodes an immunogenic 18-kDa membrane-associated protein; however, antibodies to this protein are not detected in all infected mice.

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