Serological and Molecular Characteristics of the First Korean Case of Echinococcus multilocularis

In December 2011, we reported an autochthonous case of Echinococcus multilocularis infection in a 42-year-old woman in Korea. The diagnosis was based on histopathological findings of the surgically resected liver cyst. In the present study, we evaluated the serological and molecular characteristics of this Korean E. multilocularis case. The patient's serum strongly reacted with affinity-purified native Em18 and recombinant Em18 antigens (specific for E. multilocularis) but negative for recombinant antigen B8/1 (reactive for Echinococcus granulosus). In immunoaffinity chromatography, the serum also strongly reacted with E. multilocularis and only weakly positive for E. granulosus. We determined the whole nucleotide sequence of cox1 (1,608 bp) using the paraffin-embedded cystic tissue which was compared with E. multilocularis isolates from China, Japan, Kazakhstan, Austria, France, and Slovakia. The Korean case showed 99.8-99.9% similarity with isolates from Asia (the highest similarity with an isolate from Sichuan, China), whereas the similarity with European isolates ranged from 99.5 to 99.6%.

[Comparison and evaluation of Echinococcus granulosus protoscoleces excretory/secretory proteins in PBS complemented with glucose, DMEM and RPMI culture media]

Preparation of proper antigens is an important issue in serology of hydatidosis. Investigators have been able to obtain excretory/secretory antigens [E/S Ags] by short-term culture of protoscoleces in a couple of culture media. However, no data are available about production rate of such antigens in different culture media. The present study was carried out to evaluate the production of E/S Ags [proteins] in PBS complemented with glucose, DMEM and RPMI culture media. To obtain E/S proteins, protoscoleces of echinococcus were cultured in PBS complemented with 10% glucose, RPMI and DMEM for 72 hours. Proteins secreted in culture media were concentrated and assayed. To characterize different components, proteins were electrophoresed on SDSPAGE. Data were analyzed using One-way ANOVA and Tukey HSD tests. The mean concentration of E/S proteins in PBS medium in 24 hours of culture was significantly higher than DMEM and RPMI [P<0.05]. However, such a difference was not observed between E/S proteins in DMEM and RPMI media. E/S proteins obtained from PBS medium were separated into 12 major bands and the two other media into 14 major bands within a range of molecular masses of 16 to 67 kDa. PBS complemented with glucose is more appropriate than the two other media for E/S proteins production. The best time to obtain E/S proteins is the first 48 hours of culture

Dot-Elisa for evaluation of hydatid cyst wall, protoscoleces and hydatid cyst fluid antigens in the serodiagnosis of cystic echinococcosis / Dot-Elisa para avaliação da parede do cisto hidático, protoscoleces e antígenos do fluido do cisto hidático no sorodiagnóstico da equinococose cística

The aim of the present study is to evaluate cyst wall and protoscolex as an alternate source of antigen in serodiagnosis of cystic echinococcosis (CE). A total of 90 blood samples, 30 each of confirmed CE cases, disease controls and healthy controls were collected. Dot-ELISA using cyst wall, protoscolex and cyst fluid were used to demonstrate anti-hydatid antibodies. The sensitivity of Dot-ELISA using cyst wall, protoscolex and cyst fluid was 96.66 percent, 86.66 percent and 93.33 percent respectively and the specificity of the assay was 70 percent for Dot-ELISA using cyst fluid, protoscolex and cyst wall antigens. Results of the present study show that cyst wall and protoscolex can also be an useful source of antigen in detection of hydatid antibodies in the serodiagnosis of CE.

O propósito do presente trabalho é avaliar a parede cística e protoscolex como fontes alternativas de antígeno no sorodiagnóstico de equinococose cística (CE). De um total de 90 amostras de sangue, foram coletadas 30 de casos CE confirmados, 30 de controles de doença e 30 controles saudáveis. Dot-Elisa usando parede cística, protoscolex e fluido cístico foi utilizada para demonstrar anticorpos anti-hidáticos. A sensitividade de Dot-Elisa usando parede cística, protoscolex e fluido cístico foi de: 96,66 por cento, 86,66 por cento e 93,33 por cento respectivamente e a especificidade do ensaio de 70 por cento para Dot-Elisa usando fluido cístico, protoscolex e antígeno da parede cística. Resultados do presente estudo mostram que parede cística e protoscolex podem ser fontes úteis de antígeno na detecção de anticorpos hidáticos no sorodiagnóstico do CE.

Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus

Hydatid cyst fluid (HCF), somatic antigens (S-Ag) and excretory-secretory products (ES-Ag) of Echinococcus granulosus protoscoleces are used as the main antigenic sources for immunodiagnosis of human and dog echinococcosis. In order to determine their non-shared as well as their shared antigenic components, these extracts were studied by ELISA-inhibition and immunoblot-inhibition. Assays were carried out using homologous rabbit polyclonal antisera, human sera from individuals with surgically confirmed hydatidosis, and sera from dogs naturally infected with E. granulosus. High levels of cross-reactivity were observed for all antigenic extracts, but especially for ES-Ag and S-Ag. Canine antibodies evidenced lesser avidity for their specific antigens than antibodies from human origin. The major antigenic components shared by HCF, S-Ag, and ES-Ag have apparent molecular masses of 4-6, 20-24, 52, 80, and 100-104 kDa, including doublets of 41/45, 54/57, and 65/68 kDa. Non-shared polypeptides of each antigenic extract of E. granulosus were identified, having apparent masses of 108 and 78 kDa for HCF, of 124, 94, 83, and 75 kDa for S-Ag, and of 89, 66, 42, 39, 37, and 35 kDa for ES-Ag.

Detección de inmunoglobulinas G y E específicas para E. granulosus en hidatidosis humana / Detection of G and E specific immunoglobulins for E. granulosus in human hydatidosis

Con la finalidad de conocer la utilidad clínica de la detección de anticuerpos específicos al Echinoccus granolusos de tipo inmunoglobulinas (Ig) G y E, se estudiaron 48 pacientes portadores de hidatidosis y 110 controles. Se realizó Enzima Inmuno Ensayo (ELISA) indirecto homogéneo para Ig-G y ELISA reversa para Ig-E. Se encontró una sensibilidad de 81 por ciento para IgG y de 62,5 por ciento para IgE, con una especificidad para Ig-G de 89 por ciento y para Ig-E de 96 por ciento . Los hallazgos evidencian la utilidad clínica de la detección de ambos anticuerpos específicos para el diagnóstico de la hidatidosis humana; Ig-G como primera aproximación diagnóstica y pàra estudios seroepidemiológicos e Ig-E en primoinfección, recidiva y reinfección postcirugía

role of immunoglobulins and complements in immunity against hydatis disease in Libyans

The serum immunoglobulins [IgG. IgM, IgA, IgE] and [C3, C4] levels were investigated in 45 [17 male and 28 female] Libyan patients with hydatid disease confirmed [age: 19-70 years; mean age: 44 years]. Twenty six healthy Libyans, age and sex matched, were also included in the study as control subjects. Of these 45 patients 34 had hepatic [76%]. 5 had pulmonary [11%] and 6 had multiple [hepatic, pulmonary and intestinal] [13%] lesions. The study showed that patients with hydatid disease had significantly higher total IgG [P<0.05], Igm [P<0.02] and IgE [P<0.01] levels as compared to control. Further statistical analyses for linear, exponential and geometrical correlations revealed that IgG had significant linear correlation with anti-echinococcus antibody titre [AEAT] [r=0.9214, t=5.7392, p<0.01]. Interestingly, IgE exhibited more significant exponential correlation [r=0.9309, t=5.7011, p<0.01] than geometrical correlation [r=0.8402], t=3.4643, p<0.02] with AEAT. These observations suggested than the host's immune response did occur concomitant with the existence and fluorishment of the hydatid cyst. The various possibilities as to how this can happen have been discussed

Simple and rapid-micro-indirect hemagglutination test for hydatid disease

A rapid micro-indirect haemagglutination technique using glutaraldehyde fixed-antigen coated lyophilized sheep red blood cells [GFACL-SRBCs] has been described. The test is rapid, stable, economical and shows similar results to those of freshly prepared SRBCs

Ig-G específica para E. granulosus / Specific Ig-G for E. granulosus

Nosotros desarrollamos un enzimo inmuno ensayo para detectar anticuerpos Ig-G específicos para Equinococcus Granulosus (EG) en pacientes portadores de Hidatidosis Humana confirmados quirúrgicamente. 13/18 (72.2) con localización pulmonar fueron positivos y 8/9 (88.9%) con localización hepática fueron positivos, lo que dió un total de 75.8%. Y en las formas complicada y no complicada dió un 83.3% y 100% respectivamente. A comparar con DD5 en 18 pacientes, sólo 8 (44.0%) fueron positivos para DD5 y 15 (83.0) fueron positivos para IgG (p < 0.05). Estos resultados muestran que la detección de IgG específica para EG tiene un alto rendimiento diagnóstico en la Enfermedad Hidatídica