Molecular Cloning, Expression, and Serological Evaluation of an 8-Kilodalton Subunit of Antigen B from
            <i>Echinococcus multilocularis</i>

Mamuti, Wulamu; Yamasaki, Hiroshi; Sako, Yasuhito; Nakao, Minoru; Xiao, Ning; Nakaya, Kazuhiro; Sato, Naoki; Vuitton, Dominique A.; Piarroux, Renaud; Lightowlers, Marshall W.; Craig, Philip S.; Itō, Akira

doi:10.1128/jcm.42.3.1082-1088.2004

Cited by 48 publications

(45 citation statements)

References 30 publications

(34 reference statements)

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“…The concentrations of rAgB used in the elaboration of each test (much higher in the ICT) could also have contributed to the observed differences in case detection. Nine (47.4%) of 19 AE patients' sera showed a positive result by ICT, and this result was concordant with the previous reports showing that 40 to 85% of AE patient sera reacted with rAgB by ELISA (17,18); however, since these previous studies were performed in arctic/subarctic areas where AE and CE are coendemic, this cross-reaction will not be an issue if we use the ICT in Peru, which is located in a subtropical area where infection by E. multilocularis has not been reported. In addition, in our study, sera from healthy subjects and patients with other parasitic infections except AE showed a negative result in the ICT, which indicated that this ICT was a highly specific test.…”

Section: Discussionsupporting

confidence: 80%

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Evaluation of a New Immunochromatographic Test Using Recombinant Antigen B8/1 for Diagnosis of Cystic Echinococcosis

et al. 2015

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i Diagnosis of cystic echinococcosis (CE) is based on the identification of the cyst(s) by imaging, using immunodiagnostic tests mainly as complementary tools in clinical settings. Among the antigens used for immunodiagnosis, previous studies described a good performance of the recombinant antigen B8/1 (rAgB) in an enzyme-linked immunosorbent assay (ELISA) format; however, in remote parts of areas where the disease is endemic, the implementation of an ELISA is difficult, so a more simple, rapid, and reliable method such as the immunochromatographic test (ICT) is required. In this study, using a set of 50 serum samples from patients with surgically confirmed CE, we compared the performance of an ICT and that of an ELISA using the rAgB. The overall sensitivities of ICT and ELISA were not statistically different (78% versus 72%; P ‫؍‬ 0.36). The overall agreement between both tests was moderate ( ‫؍‬ 0.41; P < 0.01). Concordance between ICT and ELISA was substantial or almost perfect for patients with liver involvement ( ‫؍‬ 0.65; P < 0.001) and patients with more than one hydatid cyst ( ‫؍‬ 0.82; P < 0.001), respectively. Moreover, specificity analysis using a total of 88 serum samples from healthy individuals (n ‫؍‬ 20) and patients (n ‫؍‬ 68) with other parasitic infections revealed that ICT had a specificity of 89.8%. ICT and ELISA had similar performance for the detection of specific antibodies to E. granulosus, and ICT had a high specificity, opening the possibility of using ICT as a screening tool in rural settings. Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of the dog tapeworm Echinococcus granulosus. This zoonosis has a worldwide distribution, being considered a public health problem in areas dedicated to the raising of livestock where CE is endemic (1). In these areas, risk factors such as access of dogs to contaminated viscera and close contact between infected dogs and humans facilitate and maintain the transmission of the disease (2, 3). Humans are infected by the accidental ingestion of the tapeworm eggs, which can develop to the larval stage (hydatid cyst) in any internal organ after several years. The organs more frequently involved are the liver and the lungs, representing ϳ70% and ϳ20% of cases, respectively (4). Diagnosis of CE is based on the identification of the hydatid cyst(s) by imaging methods (e.g., abdominal ultrasound, chest X ray, or computed tomography) (5). Immunodiagnostic tools are of use in clinical settings as a complementary diagnostic tool and have quite variable performance, which depends on the antigen or technique used and is affected by certain disease characteristics, such as cyst location and presence of cyst rupture or aggregated bacterial infection (6-9).Among the antigens used for immunodiagnostic, hydatid cyst fluid (HCF) has been the one most widely used. Antibody-detecting assays using HCF report sensitivities between 75% and 95% with poor specificity and frequent cross-reactions (5, 9-11). More recently, synthetic peptides or recombi...

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Section: Discussionsupporting

confidence: 80%

“…The recombinant 8-kDa subunit of AgB8/1 (rAgB) was expressed in a bacterial system as described previously (17) with some modifications. Briefly, a DNA fragment encoding the AgB8/1 was amplified by PCR with the primers 5=-GGGAATTCG ATGATGGTTACTCGACG-3= and 5=-TTGGATCCTTACTTTGAATCA TCATCTTT-3=.…”

Section: Serum Samplesmentioning

confidence: 99%

Evaluation of a New Immunochromatographic Test Using Recombinant Antigen B8/1 for Diagnosis of Cystic Echinococcosis

et al. 2015

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“…Mamuti and colleagues [47] used a recombinant AgB8/1, reporting sensitivities of 88.0% (95% CI 75.7-95.5%) for CE and 37.8% (95% CI 23.7-53.5%) for AE. In another study by Li and colleagues [48], a recombinant AgB subunit 8, basically prepared as described by Mamuti and colleagues [47], reached sensitivities of 77.6% (95% CI 77.6-82.7%) for CE and 85.0% (95% CI 78.8-89.9%) for AE; in terms of AE, this is significantly more sensitive than that reached by Mamuti and colleagues with the recombinant AgB8/1, but not more sensitive than that found in our study using native AgB ([99.9%; 95% CI 94.3-100%). Sensitivities for CE did not differ significantly between all three studies and were comparable with the results of a study by Gihan and colleagues [49] where a sensitivity of 82.5% (95% CI 73.2-95.8%) was reported.…”

Section: Discussionmentioning

confidence: 99%

Serological diagnosis of echinococcosis: the diagnostic potential of native antigens

et al. 2011

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PURPOSE: Human alveolar (AE) and cystic echinococcosis (CE) caused by the metacestode stages of Echinococcus multilocularis and E. granulosus, respectively, lack pathognomonic clinical signs. Diagnosis therefore relies on the results of imaging and serological studies. The primary goal of this study was to evaluate the efficacy of several easy-to-produce crude or partially purified E. granulosus and E. multilocularis metacestode-derived antigens as tools for the serological diagnosis and differential diagnosis of patients suspicious for AE or CE. METHODS: The sera of 51 treatment-naïve AE and 32 CE patients, 98 Swiss blood donors and 38 patients who were initially suspicious for echinococcosis but suffering from various other liver diseases (e.g., liver neoplasia, etc.) were analysed. RESULTS: According to the results of enzyme-linked immunosorbent assays (ELISA), metacestode-derived antigens of E. granulosus had sensitivities varying from 81 to 97% and >99.9% for the diagnosis of CE and AE, respectively. Antigens derived from E. multilocularis metacestodes had sensitivities ranging from 84 to 91% and >99.9% for the diagnosis of CE and AE, respectively. Specificities ranged from 92 to >99.9%. Post-test probabilities for the differential diagnosis of AE from liver neoplasias, CE from cystic liver lesions, and screening for AE in Switzerland were around 95, 86 and 2.2%, respectively. Cross-reactions with antibodies in sera of patients with other parasitic affections (fasciolosis, schistosomosis, amebosis, cysticercosis, and filarioses) did occur at variable frequencies, but could be eliminated through the use of confirmatory testing. CONCLUSIONS: Different metacestode-derived antigens of E. granulosus and E. multilocularis are valuable, widely accessible, and cost-efficient tools for the serological diagnosis of echinococcosis. However, confirmatory testing is necessary, due to the lack of species specificity and the occurrence of cross-reactions to other helminthic diseases. Abstract Purpose Human alveolar (AE) and cystic echinococcosis (CE) caused by the metacestode stages of Echinococcus multilocularis and E. granulosus, respectively, lack pathognomonic clinical signs. Diagnosis therefore relies on the results of imaging and serological studies. The primary goal of this study was to evaluate the efficacy of several easy-to-produce crude or partially purified E. granulosus and E. multilocularis metacestode-derived antigens as tools for the serological diagnosis and differential diagnosis of patients suspicious for AE or CE. Methods The sera of 51 treatment-naïve AE and 32 CE patients, 98 Swiss blood donors and 38 patients who were initially suspicious for echinococcosis but suffering from various other liver diseases (e.g., liver neoplasia, etc.) were analysed. Results According to the results of enzyme-linked immunosorbent assays (ELISA), metacestode-derived antigens of E. granulosus had sensitivities varying from 81 to 97% and [99.9% for the diagnosis of CE and AE, respectively. Antigens derived from...

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“…Each diagnosis of AE and CE had been carried out by imaging techniques, clinical findings, histological observations (if feasible), and/or serology of IB with recombinant Em18 (16) or EmAgB8/1 (13). As shown in Table 1, 47 AE and 2 CE patient sera were determined to be positive by ICT, and none of sera from healthy persons showed positive reactions; thus, the sensitivity and specificity of ICT were 94.0 and 95.4%, respectively.…”

Section: Alveolar Echinococcosis (Ae) Caused By the Larval Stage Ofmentioning

confidence: 99%

Development of an Immunochromatographic Test To Detect Antibodies against Recombinant Em18 for Diagnosis of Alveolar Echinococcosis

Sako

Fukuda²,

Kobayashi³

et al. 2009

J Clin Microbiol

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An immunochromatographic test (ICT) for the rapid detection of antibodies to Echinococcus multilocularis was developed. The ICT showed a sensitivity of 94% and a specificity of 95.4%. High degrees of agreement were observed between the ICT and an enzyme-linked immunosorbent assay ( ‫؍‬ 0.93) and between the ICT and immunoblot analysis ( ‫؍‬ 0.97). It is expected that the ICT developed in this study will be useful for the serodiagnosis of alveolar echinococcosis.

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Molecular Cloning, Expression, and Serological Evaluation of an 8-Kilodalton Subunit of Antigen B from Echinococcus multilocularis

Cited by 48 publications

References 30 publications

Evaluation of a New Immunochromatographic Test Using Recombinant Antigen B8/1 for Diagnosis of Cystic Echinococcosis

Evaluation of a New Immunochromatographic Test Using Recombinant Antigen B8/1 for Diagnosis of Cystic Echinococcosis

Serological diagnosis of echinococcosis: the diagnostic potential of native antigens

Development of an Immunochromatographic Test To Detect Antibodies against Recombinant Em18 for Diagnosis of Alveolar Echinococcosis

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