Diagnostic methods for differentiation of Entamoeba histolytica and Entamoeba dispar in carriers: performance and clinical implications in a non-endemic setting

Unpreserved faecal samples, suspected to contain Entamoeba histolytica/Entamoeba dispar cysts or trophozoites on the basis of microscopic examination, and serum samples from 416 patients were collected in a prospective study to determine whether stool antigen assays and detection of antibodies in serum are reliable methods to distinguish between carriers of E. histolytica and E. dispar in comparison to the reference test: real-time PCR. In 283 patients (68%) DNA of E. histolytica or E. dispar was amplified by real-time PCR: 6 patients with amoebic colitis (2%), 19 carriers of E. histolytica (6.7%), and 258 carriers of E. dispar (91.2%). In 133 patients (31%) no DNA of E. histolytica or E. dispar could be amplified in the stool samples. This patient group was used as control for the evaluation of diagnostic tests. Using real-time PCR as a reference test, the sensitivity and specificity of (1) the Entamoeba test for the diagnosis of E. histolytica/E. dispar carrier were 59% and 98%, (2) E. histolytica II for the diagnosis of E. histolytica carrier was 71% and 100%, and (3) serology for the diagnosis of E. histolytica infection was 83.3% and 95.2%, respectively. Applied to carriers that did not originate from an endemic country the sensitivity of serology for E. histolytica infection was 90% and specificity was 98.8%. In comparison to real-time PCR the performances of Entamoeba test and E. histolytica II lacked sensitivity for a reliable diagnosis of E. histolytica/E. dispar infection in a non-endemic setting. In carriers of E. histolytica/E. dispar from non-endemic countries the high specificity of serology can be used to establish the diagnosis of E. histolytica infection if antibodies are present.     

Effectiveness of latex agglutination slide test in the diagnosis of imported invasive amoebiasis in the emergency department

We compared a latex agglutination test (LAT) with enzyme-linked immunosorbent assay and indirect hemagglutination assay in the diagnosis of invasive amoebiasis. A retrospective biological records review has included 639 patients for whom these three serological tests were performed. The sensitivity of the LAT was 97.8% and the specificity was 97%.     

Differential Induction of Immunoregulatory Circuits of Phagocytic Cells by Gal/Gal NAc Lectin from Pathogenic and Nonpathogenic <Emphasis Type="Italic">Entamoeba</Emphasis>

INTRODUCTION: Amoebic liver abscess (ALA) is an acute inflammatory disease caused by Entamoeba histolytica. MATERIALS AND METHODS: The key player in the pathogenesis and immunoprotection is the Gal/GalNAc inhibitable lectin, possessed by both pathogenic (Entamoeba histolytica) and nonpathogenic (Entamoeba dispar) strains, but only E. histolytica infection is associated with an acute inflammation and subsequently the disease. RESULTS AND DISCUSSION: The stimulation with the lectin induces the secretion of various proinflammatory cytokines/chemokines from intestinal epithelial cells. The differential induction of cytokine/chemokine network by the two strains can further regulate the immunoregulatory functions of the immune cells (monocytes and neutrophils) of the host. The soluble levels of IL-1beta, IL-6, IL-8, IL-10, MIP-1alpha, MCP-1, RANTES, GRO-alpha, GMCSF were quantitated to be significantly higher in pathogenic lectin-induced conditioned media (LCM) compared to nonpathogenic LCM (NP-LCM). The monocytes from ALA patients responded to the presence of pathogenic-LCM (P-LCM) by lowering of intracellular Ca(2+) (which was still higher than control). The proinflammatory MCP-1, GRO-alpha, and GMCSF levels in the monocytes were recorded both (quantitatively and mRNA quantitation) to be tremendously higher along with their respective receptors, with P-LCM compared to NP-LCM. A significant increase in the reactive oxygen intermediates and chemotactic index (CI) was observed in the monocytes when treated with P-LCM. Similarly, in neutrophils of ALA patients, an increase in intracellular Ca(2+), ROS and chemotaxis was observed with P-LCM. OBJECTIVES: The study is a step towards understanding the mechanism of immunopathogenesis of amoebiasis, on one hand, and points to the central role of cytokine/chemokine network in the process, on the other hand.     

药物牙膏对齿龈内阿米巴的体外抑制作用

目的 研究药物牙膏对齿龈内阿米巴的体外抑制作用.方法 从牙龈炎患者口腔中取得齿龈内阿米巴体外培养,加入不同浓度的牙膏A、B、C和高露洁48h后,显微镜下观察齿龈内阿米巴形态和数目的 变化.结果 牙膏A、B、C和高露洁均可以抑制口腔齿龈内阿米巴,其IC50值分别为3.58、3.93、3.72和3.56mg/ml.结论 4...     

上海市综合性医院腹泻患者中溶组织内阿米巴感染现状调查

目的 了解综合性医院腹泻患者溶组织内阿米巴感染现状,为防治工作提供科学依据。方法 选择上海市3所综合性医院肠道门诊,采集门诊腹泻患者新鲜粪便和血清,分别采用生理盐水涂片法和碘液染色法、免疫层析法、ELISA法进行检测,以了解腹泻患者溶组织内阿米巴感染状况,并对感染者特征进行分析。结果 检测腹泻患者粪便样本 1 015份, 检出溶组织内阿米巴原虫病原学阳性36份, 总阳性率为3.55%。3所医院腹泻患者病原学阳性率间差异无统计学意义（P > 0.05）,溶组织内阿米巴阳性者性别、年龄、职业和文化程度分布差异均无统计学意义（P均 > 0.05）,脓血便中溶组织内阿米巴阳性率显著高于稀便和水样便（P均 < 0.01）。7-9月为发病高峰。88.90%的阳性者有腹痛,75.00%和22.23%的阳性者粪便查见白细胞和红细胞。试剂条法检测溶组织内阿米巴粪抗原阳性率为8.18%（83/1 015）,ELISA 法检测溶组织内阿米巴IgG抗体阳性率为7.12%（48/675）。结论 夏秋季是溶组织内阿米巴感染高发季节,应加强监测;脓血便中溶组织内阿米巴检出阳性率较高,联合应用多种检测手段能提高检出率。     

Invasive amebiasis in men who have sex with men, Australia

Entamoeba histolytica is a pathogenic ameba that has recently been recognized as an emerging pathogen in men who have sex with men (MSM) in Asia-Pacific countries where it is not endemic, i.e., Japan, Taiwan, and Republic of Korea. We report locally acquired invasive amebiasis in Sydney, Australia, exclusively in MSM.     

齿龈内阿米巴的超微结构与溶酶体酶细胞化学研究

目的　研究齿龈内阿米巴的细胞器与功能 ,探讨虫体与宿主细胞的关系。方法　用透射电镜结合电镜酶细胞化学技术观察虫体细胞超微结构 ;将虫体与口腔上皮细胞孵育 ,观察虫体对上皮细胞的粘附。结果　虫体及其伪足呈多形态 ;胞质内细胞器主要为泡状和管状的溶酶体、一些滑面内质网 ,以及丰富的糖原颗粒 ;溶酶体标志酶CMPase显示溶酶体酶丰富 ;可见虫体粘附于上皮细胞表面和小伪足伸入上皮细胞微绒毛间。结论　虫体的溶酶体水解酶消化作用及虫体伪足的机械作用对宿主牙周上皮组织的损伤 ,是虫体的致病机制之一。     

齿龈内阿米巴形态学的观察

目的 :观察齿龈内阿米巴滋养体的形态。方法 :从口腔疾病患者牙周组织或龈隙挑取渗出物 ,生理盐水涂片后镜下观察其形态并测量大小。结果 :齿龈内阿米巴滋养体活体大小平均为 14 .93× 12 .4 2μm ,形态呈圆形、长椭圆形及不规则葫芦形等。伪足透明 ,呈指状、舌状 ,球形及草帽形等不规则形态 ,其伪足伸缩变化较大 ,运动无一定方向。与溶组织内阿米巴滋养体形态有很多相似之处。结论 :全面观察了其运动方式及其形态变化特点 ,因取材方便 ,建议在寄生虫学教学实习中代替溶组织内阿米巴滋养体 ,观察活体阿米巴运动     

用限制性内切酶分析溶组织内阿米巴的致病性

用SH-3、SH-5、SH-6、SH-7和SH-85株溶组织内阿米巴的DNA增殖35个周期,将其基因DNA用内切酶HinfⅠ和EcoT22Ⅰ进行消化后,作琼脂糖凝胶电泳分析,显示,5株虫株DNA经35个循环周期后产生531-bp的产物;经HinfⅠ消化后,SH-3、SH-5、SH-6和SH-7的基因DNA产生3个相同的片段,而显示其均与非致病性虫株SAW142的电泳谐完全相同;而SH-8基因DNA的电泳谱与致病性虫株SAW408的电泳谱一致,而用EcoT22Ⅰ消化结果也显示SH-8的图谱与致病性虫株SAW408的相同,证实了从包囊携带者和有发热、腹泻而无脓血便患者粪便中分离的虫株均属于非致病性虫株,从有发热、腹泻、脓血便的急性阿米巴痢疾患者粪便中分离到的虫株属于致病性虫株,均与酶株群分析相一致。提示,应用多聚酶链反应和限制性内切酶消化基因DNA来检测溶组织内阿米巴的基因型是十分有意义的。     

Antibodies to the serine rich Entamoeba histolytica protein (SREHP) prevent amoebic liver abscess in severe combined immunodeficient (SCID) mice

Amoebic liver abscess caused by Entamoeba histolytica is a major cause of morbidity and mortality worldwide. We used mice with severe combined immunodeficiency (SCID mice) to study the role of antibody in protection from amoebic liver abscess, and to identify protective antigens of E. histolytica. Antisera to recombinant versions of two major surface antigens of E. histolytica, the serine rich E. histolytica protein (SREHP) and the 170 kDa adhesin were used in this study. We found that 100% of SCID mice passively immunized with antiserum to the recombinant SREHP molecule were protected from developing amoebic liver abscess after intrahepatic challenge with virulent E. histolytica trophozoites. In contrast, preimmune serum, antiserum to a portion of the 170 kDa adhesin, and antiserum to the trpE fusion partner of SREHP did not protect SCID mice from amoebic liver abscess. Our study demonstrates that antibodies to a recombinant version of the amoebic SREHP molecule can protect against amoebic liver abscess, and suggest the recombinant SREHP molecule should be considered as a possible vaccine candidate to prevent amoebic liver abscess.