Liu Guangxue, Liang Panhong, Zhang Shaohua, Guo Aijiang, Wang Lijie, Zheng Yadong, Luo Xuenong
State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Public Health, Ministry of Agriculture/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Public Health, Ministry of Agriculture/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Acta Trop. 2017 Jul;171:80-85. doi: 10.1016/j.actatropica.2017.03.026. Epub 2017 Mar 28.
Cysticercosis, caused by metacestodes of Taenia solium, has a significant soci-economic impact and is of considerable importance in public health. However, there are no specific diagnostic antigens to distinguish between T. solim and Taenia hydatigena. In the present study, cAMP-dependent protein kinase regulatory subunit (TsPKA-r), an excretory/secretary (ES) antigen of T. solium, was used to establish a specific and sensitive diagnostic tool for detection of porcine cysticercosis. The full-length sequence encoding TsPKA-r was amplified by PCR, sequenced and then identified by bioinformatics. The fusion protein with 6×His-tags was expressed in E. coli, purified by Ni Sepharose™ 6 Fast Flow and used to test reactionogenicity by immunoblotting. TsPKA-r based indirect enzyme-linked immunosorbent assays (iELISA) showed good performance in recognition of sera of pigs experimentally infected with T. solium metacestodes, with 93.88% sensitivity and 96.40% specificity. There were no cross-reactions against the sera from pigs experimentally infected with T. hydatigena, Toxoplasma gondii or Trichinella spiralis. These results indicate that the TsPKA-r is a promising immunodiagnostic antigen for detection of porcine cysticercosis.
由猪带绦虫的中绦期幼虫引起的囊尾蚴病具有重大的社会经济影响,在公共卫生领域相当重要。然而,目前尚无区分猪带绦虫和泡状带绦虫的特异性诊断抗原。在本研究中,猪带绦虫的排泄/分泌(ES)抗原——环磷酸腺苷(cAMP)依赖性蛋白激酶调节亚基(TsPKA-r),被用于建立一种检测猪囊尾蚴病的特异性和敏感性诊断工具。通过聚合酶链反应(PCR)扩增编码TsPKA-r的全长序列,进行测序,然后通过生物信息学进行鉴定。带有6×组氨酸标签的融合蛋白在大肠杆菌中表达,经镍琼脂糖凝胶6快速流(Ni Sepharose™ 6 Fast Flow)纯化,并通过免疫印迹法检测其反应原性。基于TsPKA-r的间接酶联免疫吸附测定(iELISA)在识别经猪带绦虫中绦期幼虫实验感染猪的血清方面表现良好,敏感性为93.88%,特异性为96.40%。对经泡状带绦虫、刚地弓形虫或旋毛虫实验感染猪的血清无交叉反应。这些结果表明,TsPKA-r是一种很有前景的用于检测猪囊尾蚴病的免疫诊断抗原。