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TsPKA-r:一种用于检测猪囊尾蚴病的潜在免疫诊断抗原。

TsPKA-r: a potential immunodiagnostic antigen for the detection of porcine cysticercosis.

作者信息

Liu Guangxue, Liang Panhong, Zhang Shaohua, Guo Aijiang, Wang Lijie, Zheng Yadong, Luo Xuenong

机构信息

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.

出版信息

Acta Trop. 2017 Jul;171:80-85. doi: 10.1016/j.actatropica.2017.03.026. Epub 2017 Mar 28.

Abstract

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.

摘要

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。

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