TsPKA-r as a potential immunodiagnostic antigen for the detection of porcine cysticercosis
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- 英文论文题名：TsPKA-r as a potential immunodiagnostic antigen for the detection of porcine cysticercosis
- 论文作者：Guangxue Liu ; Panhong Liang ; Aijiang Guo ; Shaohua Zhang ; Lijie Wang ; Xuenong Luo
- 英文论文作者：Guangxue Liu ; Panhong Liang ; Aijiang Guo ; Shaohua Zhang ; Lijie Wang ; Xuenong Luo ; 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
- 年：2016
- 作者机构：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;
- 英文论文关键词：TsPAK-r ; T.solium ; ELISA ; immunodiagnostic antigens
- 会议召开时间：2016-08-08
- 会议录名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会论文摘要集
- 英文会议录名称：The Abstracts of the Tenth Symposium for Yong Scientists of China Society of Parasitology
- 语种：英文
- 分类号：S858.28
- 学会代码：JISC
- 会议名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会
- 会议地点：中国四川成都
- 主办单位：中国动物学会寄生虫学专业青年委员会
- 学会名称：中国动物学会寄生虫学专业委员会
- 页数：1
- 文件大小：136k
- 原文格式：D
- 会议级别：全国
摘要
Background: Cysticercosis, caused by the metacestode larval stage of Taenia solium, has a significant socio-economic impact and also is of considerable importance in the public health. However, there is no known specific diagnostic antigen to detect for porcine cysticercosis. The present study identified an ES antigen of T.solium and used the recombinant antigen to establish specific and sensitive diagnostic tool for detection of porcine cysticercosis. Methods: c AMP-dependent protein kinase regulatory subunit(TsPKA-r) gene was searched through database of previously published T.solium genome and transcriptome data. The full-length sequence encoding TsPKA-r was amplified from adult worm c DNA by using 3'5'-RACE-PCR and subsequently cloned into the prokaryotic expression vector p ET30a(+). TsPKA-r fusion-protein with his-tag was expressed in E.coli, purified by Ni SepharoseTM 6 Fast Flow and used to test the reactogenicity by immunoblotting. Furthermore, the indirect enzyme-linked immunosorbent assays(iE LISA) based on TsPKA-r was established using the serum of pigs experimentally infected with cysticercus cellulosae. Results: Analyzing deduced amino acid sequence of the TsPKA-r gene revealed that it contains the essential structural motif of PKA family. The 54 k Da recombinant protein of TsPKA-r can be recognized by serum of pigs experimentally infected with cysticercus cellulosae, which indicated that TsPKA-r could induce immune response. The iELISA based on TsPKA-r could detect specific antibodies from pigs experimentally infected cysticercus cellulosae with 98% sensitivity and 95% specificity. It showed no cross-reactions against the serum samples from pigs experimentally infected with Cysticercus tenuicollis, Toxoplasma gondii, Trichinella spiralis. Conclusion: The current results indicated that the TsPKA-r is promising immunodiagnostic antigen for developing iELISA to detect porcine cysticercosis.
Background: Cysticercosis, caused by the metacestode larval stage of Taenia solium, has a significant socio-economic impact and also is of considerable importance in the public health. However, there is no known specific diagnostic antigen to detect for porcine cysticercosis. The present study identified an ES antigen of T.solium and used the recombinant antigen to establish specific and sensitive diagnostic tool for detection of porcine cysticercosis. Methods: c AMP-dependent protein kinase regulatory subunit(TsPKA-r) gene was searched through database of previously published T.solium genome and transcriptome data. The full-length sequence encoding TsPKA-r was amplified from adult worm c DNA by using 3'5'-RACE-PCR and subsequently cloned into the prokaryotic expression vector p ET30a(+). TsPKA-r fusion-protein with his-tag was expressed in E.coli, purified by Ni SepharoseTM 6 Fast Flow and used to test the reactogenicity by immunoblotting. Furthermore, the indirect enzyme-linked immunosorbent assays(iE LISA) based on TsPKA-r was established using the serum of pigs experimentally infected with cysticercus cellulosae. Results: Analyzing deduced amino acid sequence of the TsPKA-r gene revealed that it contains the essential structural motif of PKA family. The 54 k Da recombinant protein of TsPKA-r can be recognized by serum of pigs experimentally infected with cysticercus cellulosae, which indicated that TsPKA-r could induce immune response. The iELISA based on TsPKA-r could detect specific antibodies from pigs experimentally infected cysticercus cellulosae with 98% sensitivity and 95% specificity. It showed no cross-reactions against the serum samples from pigs experimentally infected with Cysticercus tenuicollis, Toxoplasma gondii, Trichinella spiralis. Conclusion: The current results indicated that the TsPKA-r is promising immunodiagnostic antigen for developing iELISA to detect porcine cysticercosis.
Background: Cysticercosis, caused by the metacestode larval stage of Taenia solium, has a significant socio-economic impact and also is of considerable importance in the public health. However, there is no known specific diagnostic antigen to detect for porcine cysticercosis. The present study identified an ES antigen of T.solium and used the recombinant antigen to establish specific and sensitive diagnostic tool for detection of porcine cysticercosis. Methods: c AMP-dependent protein kinase regulatory subunit(TsPKA-r) gene was searched through database of previously published T.solium genome and transcriptome data. The full-length sequence encoding TsPKA-r was amplified from adult worm c DNA by using 3'5'-RACE-PCR and subsequently cloned into the prokaryotic expression vector p ET30a(+). TsPKA-r fusion-protein with his-tag was expressed in E.coli, purified by Ni SepharoseTM 6 Fast Flow and used to test the reactogenicity by immunoblotting. Furthermore, the indirect enzyme-linked immunosorbent assays(iE LISA) based on TsPKA-r was established using the serum of pigs experimentally infected with cysticercus cellulosae. Results: Analyzing deduced amino acid sequence of the TsPKA-r gene revealed that it contains the essential structural motif of PKA family. The 54 k Da recombinant protein of TsPKA-r can be recognized by serum of pigs experimentally infected with cysticercus cellulosae, which indicated that TsPKA-r could induce immune response. The iELISA based on TsPKA-r could detect specific antibodies from pigs experimentally infected cysticercus cellulosae with 98% sensitivity and 95% specificity. It showed no cross-reactions against the serum samples from pigs experimentally infected with Cysticercus tenuicollis, Toxoplasma gondii, Trichinella spiralis. Conclusion: The current results indicated that the TsPKA-r is promising immunodiagnostic antigen for developing iELISA to detect porcine cysticercosis.
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