Key Laboratory of Animal Parasitology of Ministry of Agriculture and Rural Affairs, Laboratory of Quality and Safety Risk Assessment for Animal Products on Biohazards (Shanghai) of Ministry of Agriculture and Rural Affairs, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, PR China.
State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Clinical Medical Research Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, PR China.
Microb Pathog. 2023 Jun;179:106110. doi: 10.1016/j.micpath.2023.106110. Epub 2023 Apr 13.
Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the metacestode larva of Echinococcus granulosus. In this study, two-dimensional gel electrophoresis (2-DE) coupled with immunoblot analysis revealed that E. granulosus severin and 14-3-3zeta proteins (named EgSeverin and Eg14-3-3zeta, respectively) might be two potential biomarkers for serological diagnosis of echinococcosis. The recombinant EgSeverin (rEgSeverin, 45 kDa) and Eg14-3-3zeta (rEg14-3-3zeta, 35 kDa) were administered subcutaneously to BALB/c mice to obtain polyclonal antibodies for immunofluorescence analyses (IFAs). And IFAs showed that both proteins were located on the surface of protoscoleces (PSCs). Western blotting showed that both proteins could react with sera from E. granulosus-infected sheep, dog, and mice. Indirect ELISAs (rEgSeverin- and rEg14-3-3zeta-iELISA) were developed, respectively, with sensitivities and specificities ranging from 83.33% to 100% and a coefficient of variation (CV %) of less than 10%. The rEgSeverin-iELISA showed cross-reaction with both E. granulosus and E. multilocularis, while the rEg14-3-3zeta-iELISA showed no cross-reaction with other sera except for the E. granulosus-infected ones. The field sheep sera from Xinjiang and Qinghai were analyzed using rEgSeverin-iELISA, rEg14-3-3zeta-iELISA, and a commercial kit respectively, and no significant differences were found among the three methods (p > 0.05). However, the CE positive rates in sheep sera from Qinghai were significantly higher than those from Xinjiang (p < 0.01). Overall, the results suggest that EgSeverin and Eg14-3-3zeta could be promising diagnostic antigens for E. granulosus infection.
泡球蚴病（CE）是一种由细粒棘球绦虫的幼虫引起的人畜共患寄生虫病。在这项研究中，二维凝胶电泳（2-DE）结合免疫印迹分析表明，细粒棘球蚴 severin 和 14-3-3zeta 蛋白（分别命名为 EgSeverin 和 Eg14-3-3zeta）可能是囊型包虫病血清学诊断的两个潜在生物标志物。重组 EgSeverin（rEgSeverin，45 kDa）和 Eg14-3-3zeta（rEg14-3-3zeta，35 kDa）被皮下注射到 BALB/c 小鼠中，以获得用于免疫荧光分析（IFA）的多克隆抗体。IFA 显示两种蛋白均位于原头蚴（PSCs）的表面。Western blot 显示两种蛋白均能与感染细粒棘球蚴的绵羊、狗和小鼠的血清发生反应。分别建立了间接 ELISA（rEgSeverin-和 rEg14-3-3zeta-iELISA），其灵敏度和特异性范围分别为 83.33%至 100%，变异系数（CV%）小于 10%。rEgSeverin-iELISA 与细粒棘球蚴和多房棘球绦虫均发生交叉反应，而 rEg14-3-3zeta-iELISA 除感染细粒棘球蚴的血清外，与其他血清均无交叉反应。使用 rEgSeverin-iELISA、rEg14-3-3zeta-iELISA 和商业试剂盒分别分析来自新疆和青海的野外绵羊血清，三种方法之间无显著差异（p>0.05）。然而，青海绵羊血清中的 CE 阳性率明显高于新疆（p<0.01）。总体而言，结果表明 EgSeverin 和 Eg14-3-3zeta 可能是细粒棘球蚴感染的有前途的诊断抗原。
