Evaluation of Em18-, Em16-, Antigen B-Western blots, Em2plus-ELISA and four other tests for differential serodiagnosis of alveolar and cystic echinococcosis patients in Poland

Ito, Akira; Ma, Liang; Paul, Małgorzata; Stefaniak, Jerzy; Pawłowski, Z.

doi:10.1016/s1383-5769(98)00010-5

Cited by 22 publications

(29 citation statements)

References 10 publications

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“…Serum samples from other parasitic diseases were also examined. Four, twenty, and nine cystic echinococcosis (CE) serum samples were from Japan (imported cases), Australia, and the United States, respectively, and ten serum samples obtained from patients with Taenia solium neurocysticercosis (NCC), clinically and serologically confirmed, were from the Centers for Disease Control (19)(20)(21)29). Australian CE samples all were serologically confirmed in Melbourne by both ELISA and immunoelectrophoresis against Arc 5 (32).…”

Section: Methodsmentioning

confidence: 99%

“…The sensitivity and specificity of Em18 for AE are very compatible to those of recombinant EM10 or truncated fragments thereof (18,21), raising the question as to whether EM10 and Em18 are antigenically or otherwise related. We here describe a partial amino acid sequence of the native Em18 antigen, confirming that Em18 is a fragment of the EM10 protein, and demonstrate that recombinant Em18 is highly effective in immunoassays for serodiagnosis of AE.…”

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

confidence: 99%

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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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Section: Methodsmentioning

confidence: 99%

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

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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“…Therefore, early diagnosis and treatment are crucial for the reduction of morbidity and mortality. Because imaging technology is not always available for local patients in areas of high endemicity, such as in China, because of poorly equipped medical facilities and high cost (7), serodiagnosis by ELISA or immunoblotting has been employed with specific and purified diagnostic antigens such as Em2 plus (4) and Em18 (5). Also, crude antigen extracts of E. multilocularis have often been used for primary screening in an epidemiological survey (8).…”

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confidence: 99%

Evaluation of Use of Recombinant Em18 and Affinity-Purified Em18 for Serological Differentiation of Alveolar Echinococcosis from Cystic Echinococcosis and Other Parasitic Infections

Xiao

Mamuti

Yamasaki³

et al. 2003

J Clin Microbiol

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To further evaluate recombinant Em18 antigen (rEm18) for immunodiagnosis of human alveolar echinococcosis, 208 serum samples were examined by enzyme-linked immunosorbent assay (ELISA). To comparatively assess the results of rEm18-ELISA, ELISA and immunoblot analysis with two affinity-purified native antigens were also performed with 45 selected serum samples. The results indicate that rEm18 is highly useful for serodiagnosis.

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“…However, Nirmalan and Craig (14) reported that both Em18 and Em16 antigens crossreacted with sera from CE patients. In addition, previous immunoblotting studies reported problems with band identification and indeterminate results (10,12). Differentiating Em18 and Em16 was found to be difficult, and the location of Em16 has been identified by a specific monoclonal antibody (8,9,10).…”

mentioning

confidence: 99%

“…In addition, previous immunoblotting studies reported problems with band identification and indeterminate results (10,12). Differentiating Em18 and Em16 was found to be difficult, and the location of Em16 has been identified by a specific monoclonal antibody (8,9,10). Purification of Em18 by affinity chromatography and production of a recombinant Em18 led to highly reliable, but not completely species-specific, diagnosis of echinococcosis by immunoblotting (11).…”

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confidence: 99%

Use of Two Sensitive and Specific Immunoblot Markers, Em70 and Em90, for Diagnosis of Alveolar Echinococcosis

et al. 2004

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Antibodies against Echinococcus multilocularis metacestodes were screened by immunoblotting sera from patients with alveolar echinococcosis (n ‫؍‬ 39), cystic echinococcosis (n ‫؍‬ 109), or other parasitic infections (n ‫؍‬ 66) and healthy individuals (n ‫؍‬ 32). Two antigens, approximately 70 and 90 kDa, are found to be valuable for confirmatory diagnosis, with a sensitivity and specificity of 100 and 99.51%, respectively.Human alveolar echinococcosis (AE), caused by the metacestode of Echinococcus multilocularis, is considered the most potentially lethal parasitic zoonosis in the nontropical areas of the Northern Hemisphere (1, 2, 7, 15). Imaging techniques, serological tests, and biopsy are used for the diagnosis of AE. However, radiological patterns of AE lesions are difficult to interpret and may resemble primary hepatic neoplasm or metastasis (3). Diagnosis is possible when the imaging findings are correlated with appropriate clinical and serological findings (3,20). Tests based on purified and recombinant antigens are reported to be useful in the differential diagnosis of patients infected with Echinococcus granulosus, the agent responsible for cystic echinococcosis (CE), and E. multilocularis (4,5,17,18). Immunoblotting is used as a confirmatory test for excluding cross-reactivity in positive sera or in assessment of the screening test results (6, 12).Two E. multilocularis metacestode antigens of 18 and 16 kDa (Em18 and Em16, respectively) were recommended for the specific diagnosis of AE by immunoblotting (6). Em18 was reported to be more specific and sensitive than Em16 (7) and is useful in both demonstrating active lesions (8) and evaluating the efficacy of chemotherapy (13). However, Nirmalan and Craig (14) reported that both Em18 and Em16 antigens crossreacted with sera from CE patients. In addition, previous immunoblotting studies reported problems with band identification and indeterminate results (10, 12). Differentiating Em18 and Em16 was found to be difficult, and the location of Em16 has been identified by a specific monoclonal antibody (8, 9, 10). Purification of Em18 by affinity chromatography and production of a recombinant Em18 led to highly reliable, but not completely species-specific, diagnosis of echinococcosis by immunoblotting (11). These limitations indicated the need for new confirmatory tests in the diagnosis of AE.In this study, antibodies against antigens of E. multilocularis metacestodes were screened for the presence of sensitive and specific markers for the serological diagnosis of AE by immunoblotting.Serum samples from 39 patients with AE and 109 patients with CE were included in this study. Diagnosis of AE is confirmed by histopathology (n ϭ 27), and patients with typical liver lesion morphology were determined to have advanced inoperable AE (n ϭ 12) by clinical and radiological imaging findings. Diagnosis of CE was confirmed by serology and typical imaging findings and/or surgery. To assess cross-reactivity, serum samples were selected from patients with other proven pa...

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Evaluation of Em18-, Em16-, Antigen B-Western blots, Em2plus-ELISA and four other tests for differential serodiagnosis of alveolar and cystic echinococcosis patients in Poland

Cited by 22 publications

References 10 publications

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

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

Evaluation of Use of Recombinant Em18 and Affinity-Purified Em18 for Serological Differentiation of Alveolar Echinococcosis from Cystic Echinococcosis and Other Parasitic Infections

Use of Two Sensitive and Specific Immunoblot Markers, Em70 and Em90, for Diagnosis of Alveolar Echinococcosis

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