Identification of a cDNA clone from the larval stage ofEchinococcus granulosus with homologies to theE. multilocularis antigen EM10-expressing cDNA clone

Frosch, Petra; Mühlschlegel, Fritz A.; Sygulla, Liliane; Hartmann, M.D.; Frosch, Matthias

doi:10.1007/bf00932958

Cited by 21 publications

(14 citation statements)

References 10 publications

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“…Nevertheless, all the seropositive reactions in CE patients were much weaker than in AE patients (OD medians, 0.135 versus 1.068). Similar results were obtained with antigens EM10 or II/3, described in previous studies (4,6), in which AE patients were found to have raised specific antibody to EM10 more frequently than CE patients, despite there being a protein with a high level of homology to EM10 expressed by E. granulosus metacestodes. The significant differences in activities with rEm18 or EM10 in AE and CE patients may be partially attributable to different pathological features of metacestodes of E. multilocularis and E. granulosus.…”

Section: Discussionsupporting

confidence: 89%

Specific IgG Responses to Recombinant Antigen B and Em18 in Cystic and Alveolar Echinococcosis in China

Itō

Chen

et al. 2010

Clin Vaccine Immunol

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An understanding of the correlation of the specific antibody responses and the disease phase is essential in evaluating diagnostic values of immunological tests in human echinococcosis. In this study, 422 echinococcosis patients diagnosed by ultrasonography, including 246 with cystic echinococcosis (CE), 173 with alveolar echinococcosis (AE), and 3 with dual infection, were tested for specific IgG in sera against recombinant AgB (rAgB) and recombinant Em18 (rEm18) in an enzyme-linked immunosorbent assay. As a result, rAgB-specific antibody was detected in 77.6% of CE and 86.1% of AE patients, while rEm18-specific antibody was present in 28.9% of CE and 87.3% of AE patients. Additionally, all three patients with dual infection exhibited specific antibodies responding to rAgB and rEm18. Further analysis revealed that rAgB-specific antibody was elevated in a significantly greater proportion (87.3%) of CE patients with cysts at active or transitional stages (CE1, CE2, or CE3), compared to 54.8% of other patients with cysts at an early or an inactive stage (CL or CE4 or CE5). Furthermore, rAgB-specific antibody was detected in 95.6% of CE2 cases, which was statistically greater than that (73.7%) in CE1 patients. Although rEm18-specific antibody was elevated in 28.9% of CE patients, the positive reaction was much weaker in CE than in AE cases. Serum levels and concentrations of rEm18-specific antibody were further indicated to be strongly disease phase correlated in AE patients, with positive rates of 97.4% in cases with alveolar lesions containing central necrosis and 66.7% in patients with early alveolar lesions that measured <5 cm.Humans acquire the infection of echinococcosis by accidental ingestion of eggs excreted with feces of carnivores harboring the adult worms of Echinococcus spp. The eggs hatch in the small intestine of humans, releasing the oncosphere, which migrates via the portal system into various organs and then develops into the metacestode stage. The larval parasite can establish itself in any part of the human body but most frequently does so in the liver (32). Diagnosis of human echinococcosis is primarily based on the pathognomonic features in images obtained using imaging techniques including ultrasonography, computed tomography (CT), and magnetic resonance imaging (MRI). Of these techniques, B-ultrasound is much more widely applied, as CT and MRI are too expensive and largely inaccessible in most areas where echinococcosis is endemic. Criteria for classification of cystic echinococcosis (CE) and alveolar echinococcosis (AE) have been proposed based on stage-specific ultrasound images (20,36). Briefly, on the basis of conformational features of cysts, CE lesions are differentiated into six types: CL, CE1, CE2, CE3, CE4, and CE5. The CL type refers to a cystic lesion of a parasite origin and without a clear rim, indicating the parasite is at a very early stage of development. The CE1 type describes a unilocular simple cyst with uniform anechoic content and, importantly, with a visible wall, while...

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Section: Discussionsupporting

confidence: 89%

Specific IgG Responses to Recombinant Antigen B and Em18 in Cystic and Alveolar Echinococcosis in China

Itō

Chen

et al. 2010

Clin Vaccine Immunol

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“…Despite there being a protein expressed by E. granulosus metacestodes which has a high level of homology to EM10 (98.6% identity at amino acid level) (4,6), this protein does not induce significant levels of cross-reacting antibodies to RecEm18 in CE patients except in a small minority of patients. Similar results were obtained by Felleisen and Gottstein (4), who found that sera from either AE or CE patients reacted similarly to both E. multilocularis EM10 (referred to by them as II/3) or its E. granulosus homologue and that AE patients more frequently raised antibodies to this antigen.…”

Section: Discussionmentioning

confidence: 99%

Alveolar Echinococcosis: Characterization of Diagnostic Antigen Em18 and Serological Evaluation of Recombinant Em18

Sako¹,

Nakao²,

Nakaya

et al. 2002

J Clin Microbiol

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The Echinococcus multilocularis protein Em18 is one of the most promising antigens for use in serodiagnosis of alveolar echinococcosis in human patients. Here we identify an antigenic relationship between Em18 and a 65-kDa immunodominant E. multilocularis surface protein previously identified as either EM10 or EmII/3. The NH 2 -terminal sequence of native Em18 was determined, revealing it to be a fragment of EM10. Experiments were undertaken to investigate the effect of proteinase inhibitors on the degradation of EM10 in crude extracts of E. multilocularis protoscoleces. Em18 was found to be the product of degradation of EM10 by cysteine proteinase. A recombinant Em18 (RecEm18, derived from 349 K to 508 K of EM10) was successfully expressed by using Escherichia coli expression system and then evaluated for use in serodiagnosis of alveolar echinococcosis. RecEm18 was recognized by 27 (87.1%) and 28 (90.3%) of 31 serum samples from clinically and/or pathologically confirmed alveolar echinococcosis patients by enzyme-linked immunosorbent assay and immunoblotting, respectively. Of 33 serum samples from cystic echinococcosis patients, 1 was recorded as having a weak positive reaction to RecEm18; however, none of the serum samples which were tested from neurocysticercosis patients (n ‫؍‬ 10) or healthy people (n ‫؍‬ 15) showed positive reactions. RecEm18 has the potential for use in the differential serodiagnosis of alveolar echinococcosis.

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“…The antigens chosen for this study included HCF, which, due to its ease of preparation, is the most popular antigenic source, and native AgB, the major parasite component of HCF, which is regarded as one of the most valuable E. granulosus antigens. Our panel also comprised three AgB-related antigens, namely, AgB8/1, AgB8/2, and the peptide p-176, which have been reported as highly antigenic in human infections (7,10,19). Additionally, EgMDH was also included on the basis of its availability and previously reported diagnostic value, and as an alternative to AgB-related antigens.…”

Section: Discussionmentioning

confidence: 99%

“…HCF was obtained by aseptic aspiration from either bovine or ovine fertile cyst (2). Seven HCF preparations were used in this study, HCF 1 to HCF 4 and HCF 5 to HCF 7 , from bovine and ovine origin, respectively. HCF 1 was analyzed in parallel in all participating laboratories.…”

Section: Methodsmentioning

confidence: 99%

Comparative Analysis of the Diagnostic Performance of Six Major Echinococcus granulosus Antigens Assessed in a Double-Blind, Randomized Multicenter Study

Lorenzo¹,

Ferreira

Monteiro

et al. 2005

J Clin Microbiol

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The serodiagnosis of hydatid disease is a valuable instrument for clinical diagnosis and epidemiological surveillance of high-risk populations. In the past decade a wealth of reports on the diagnostic performance of numerous antigens have been produced. However, their diagnostic value has been estimated under different conditions, using different serum collection, therefore precluding their direct comparison. Here we report an unbiased comparison of the same batch of six major E. granulosus antigens, namely, hydatid cyst fluid (HCF), native antigen B (AgB), two recombinant AgB subunits, an AgB-derived synthetic peptide, and recombinant cytosolic malate dehydrogenase from E. granulosus (EgMDH), against the same serum collection. The doubleblind analysis was performed using a standardized protocol and receiver operating characteristic (ROC) data analysis by a network of six South American laboratories. High intercenter reproducibility was attained, and the intralaboratory analysis allowed the comparative ranking of the antigen panel. HCF, AgB, and its AgB8/1 subunit exhibited equivalent diagnostic efficiencies, 81.4% ؎ 0.5%, 81.3% ؎ 0.6%, and 81.9% ؎ 2.0%, respectively; with a more favorable balance toward specificity in the case of the last antigen. The diagnostic efficiencies for the other three antigens were 76.8% ؎ 6.8%, 69.1% ؎ 2.7%, and 66.8% ؎ 2.1%, for the peptide, the AgB8/2 subunit, and the EgMDH, respectively. The study also included an analysis of batch-to-batch variation in the diagnostic performance of different HCF regional preparations. Based on these results, a suggested recommendation on the use of these antigens was drawn.

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Identification of a cDNA clone from the larval stage ofEchinococcus granulosus with homologies to theE. multilocularis antigen EM10-expressing cDNA clone

Cited by 21 publications

References 10 publications

Specific IgG Responses to Recombinant Antigen B and Em18 in Cystic and Alveolar Echinococcosis in China

Specific IgG Responses to Recombinant Antigen B and Em18 in Cystic and Alveolar Echinococcosis in China

Alveolar Echinococcosis: Characterization of Diagnostic Antigen Em18 and Serological Evaluation of Recombinant Em18

Comparative Analysis of the Diagnostic Performance of Six Major Echinococcus granulosus Antigens Assessed in a Double-Blind, Randomized Multicenter Study

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