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棘球蚴葡萄糖激酶的分子特征和血清学诊断潜力。

Molecular characterization and serodiagnostic potential of Echinococcus granulosus hexokinase.

机构信息

Institute of Pathogenic Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, 730000, Gansu, People's Republic of China.

The Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen, 518000, Guangdong, People's Republic of China.

出版信息

Parasit Vectors. 2021 Feb 8;14(1):105. doi: 10.1186/s13071-021-04606-8.

Abstract

BACKGROUND

Cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus (sensu stricto), is a life-threatening but neglected zoonosis. Glycolytic enzymes are crucial molecules for the survival and development of E. granulosus. The aim of this study was to investigate the molecular characterization, immunogenicity, tissue distribution and serodiagnostic potential of E. granulosus hexokinase (EgHK), the first key enzyme in the glycolytic pathway.

METHODS

EgHK was cloned and expressed in Escherichia coli. Specific serum antibodies were evaluated in mice immunized with recombinant EgHK (rEgHK). The location of EgHK in the larval stage of E. granulosus was determined using fluorescence immunohistochemistry, and the potential of rEgHK as a diagnostic antigen was investigated in patients with CE using indirect enzyme-linked immunosorbent assay (ELISA).

RESULTS

Recombinant EgHK could be identified in the sera of patients with CE and in mouse anti-rEgHK sera. High titers of specific immunoglobulin G were induced in mice after immunization with rEgHK. EgHK was mainly located in the tegument, suckers and hooklets of protoscoleces and in the germinal layer and laminated layer of the cyst wall. The sensitivity and specificity of the rEgHK-ELISA reached 91.3% (42/46) and 87.8% (43/49), respectively.

CONCLUSIONS

We have characterized the sequence, structure and location of EgHK and investigated the immunoreactivity, immunogenicity and serodiagnostic potential of rEgHK. Our results suggest that EgHK may be a promising candidate for the development of vaccines against E. granulosus and an effective antigen for the diagnosis of human CE.

摘要

背景

由细粒棘球绦虫(狭义)幼虫引起的包虫病是一种危及生命但被忽视的人畜共患病。糖酵解酶是细粒棘球绦虫生存和发育的关键分子。本研究旨在研究糖酵解途径中的第一个关键酶细粒棘球蚴己糖激酶(EgHK)的分子特征、免疫原性、组织分布和血清学诊断潜力。

方法

在大肠杆菌中克隆和表达 EgHK。用重组 EgHK(rEgHK)免疫的小鼠评估特异性血清抗体。用荧光免疫组织化学法确定 EgHK 在细粒棘球蚴幼虫期的位置,并在包虫病患者中用间接酶联免疫吸附试验(ELISA)研究 rEgHK 作为诊断抗原的潜力。

结果

重组 EgHK 可在包虫病患者的血清中和小鼠抗 rEgHK 血清中鉴定。rEgHK 免疫后,小鼠诱导产生高滴度的特异性免疫球蛋白 G。EgHK 主要位于原头节的皮层、吸盘和钩,以及囊壁的生发层和板层。rEgHK-ELISA 的敏感性和特异性分别达到 91.3%(42/46)和 87.8%(43/49)。

结论

我们已经对 EgHK 的序列、结构和位置进行了特征描述,并研究了 rEgHK 的免疫反应性、免疫原性和血清学诊断潜力。我们的结果表明,EgHK 可能是开发抗细粒棘球蚴疫苗的有前途的候选物,也是诊断人类包虫病的有效抗原。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e3a/7871637/ffdf688c66e3/13071_2021_4606_Fig1_HTML.jpg

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