Molecular-and Immuno-Biology of Echinococcosis

棘球蚴病和囊虫病是一种世界性的慢性难治性寄生虫病。这一国际合作项目的主要目的是为这些新出现的寄生虫病制定更好的鉴别血清诊断方案。为此，我们首先采用免疫印迹法。我们从多房棘球蚴（Em）原头节中发现了两个候选抗原成分，命名为en18和em16，作为肺泡棘球蚴病（AE）的推测血清诊断标志物。研究表明，em18对AE的特异性最强，对AE与囊性棘球蚴病、囊虫病等寄生虫病的鉴别具有重要意义。相反，em16现在被认为是Em和E.granulosus （Eg）之间的共享抗原。利用部分纯化的em18 / em16富集部分，我们建立了一种特异性优于市售Em2plus-ELISA的新ELISA方法。ELISA和免疫印迹法检测em18抗体是目前最可靠的鉴别诊断方法。利用针对em16的单克隆抗体，我们成功地建立了几个产生重组抗原（rEm16）的克隆，并发现em16的DNA序列与先前报道的EmII/3抗原相同。目前，我们正在尝试制备抗em18的单克隆抗体，并建立无em16污染的em18 - elisa用于AE的鉴别血清诊断。在建立CE和囊虫病的鉴别血清诊断基础上，我们还从Eg的囊肿液中发现了非常好的候选抗原，从猪带绦虫囊虫中部分纯化了抗原。使用来自最好大学的大量血清样本，我们评估了来自(a) CE的Eg和(b)囊虫病的Ts的新候选抗原的特异性。提示我们的AE、CE和囊虫病候选抗原都是高度可靠的，可用于流行国家的临床和流行病学研究。我们将从1997年开始在亚洲国家开展血清流行病学研究。少

Echinococcosis and cysticercosis are chronic and intractable parasitic diseases spreading all over the world. The main purpose of this international collaboration project was to establish better resolutions for differential serodiagnosis on these emerging parasitic diseases. For this purpose, we used immunoblot assay at first. We found previously undescribed, two candidate antigenic components from protoscolex of Echinococcus multilocularis (Em), designated En 18 and Em 16, as putative serodiagnostic markers unique to alveolar echinococcosis (AE). It has been evaluated that Em 18 is the most specific to AE and highly useful for differentiation of AE from other parasitic diseases including cystic echinococcosis (CE) and cysticercosis. In contrast, Em 16 is now known as shared antigen between Em and E.granulosus (Eg). Using partially purifed Em 18/Em 16 enriched fraction, we have established a new ELISA mthod with better specificity than Em2plus-ELISA,only commercially available. Our new … More ELISA and immunoblot to detect antibody against Em 18 is the most reliable for differential serodiagnosis so far. Using monoclonal antibody against Em 16, we have just succeeded in establishing several clones producing recombinant antigen (rEm16) and revealed that DNA sequence of Em 16 is the same to that of EmII/3 antigen reported previously. At this stage, we are trying to produce monoclonal antibody against Em 18 and establish Em 18-ELISA without contamination of Em 16 for differential serodiagnosis of AE.On the establishment of differential serodiagnosis of CE and cysticercosis, we also have found very good candidate antigens from cyst fluid of Eg and partially purified antigens from cysticerci of Taenia solium (Ts). Using huge number of serum samples from the best colleages, we have evaluated the specificity of new candidate antigens from (a) Eg for CE and (b) Ts for cysticercosis. It is strongly suggested that our new candidate antigens for AE,CE and cysticercosis all are highly reliable and useful for clinical and epidemiological study in endemic countries. We are going to do seroepidemiological study in Asian countries from 1997 as a new project. Less