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Cloning, expression and serodiagnostic potential of HSP70 of Taenia multiceps in sheep
Molecular and Biochemical Parasitology ( IF 1.4 ) Pub Date : 2021-07-17 , DOI: 10.1016/j.molbiopara.2021.111397
Seyma Gunyakti Kilinc 1 , Sami Simsek 2 , Figen Celik 2 , Harun Kaya Kesik 3
Affiliation  

The aim of this study was to determine the efficacy of the recombinant protein expressed by the T. multiceps HSP70 gene in the immunodiagnosis of sheep coenurosis. Specific primers were designed to amplify the gene encoding HSP70 from the whole genome sequence of T. multiceps, mRNA was isolated from C. cerebralis cysts obtained from an infected sheep’s brain, and then, cDNA was isolated. The gene region related to the designed primers was amplified by PCR, and the obtained product was cloned and expressed. The specificity of the recombinant protein purified afterwards was demonstrated by Western Blot using known positive and negative sheep sera. Additionally, the efficiency of the recombinant protein in ELISA was determined with sera collected from 1207 sheep. Finally, the sensitivity and specificity values ​of the recombinant TmHSP70 antigen in the Western Blot were 100 %, while the sensitivity and specificity rates of the ELISA were 66.6 % and 52.6 %, respectively. At the same time, 567 (46.97 %) of the 1207 analyzed serum samples were found to be positive by ELISA. In conclusion, the Western Blot technique had a higher quality than ELISA in detecting the TmHSP70 antigen for the serodiagnosis of sheep coenurosis.



中文翻译:

绵羊多头带绦虫HSP70的克隆、表达及血清诊断潜力

本研究的目的是确定由T. multiceps HSP70 基因表达的重组蛋白在绵羊胆囊病免疫诊断中的功效。设计特异性引物,从多头牛全基因组序列中扩增编码HSP70的基因,从梭菌中分离mRNA从受感染的绵羊脑中获得包囊,然后分离出 cDNA。PCR扩增与设计引物相关的基因区域,对所得产物进行克隆表达。之后纯化的重组蛋白的特异性通过Western Blot使用已知的阳性和阴性绵羊血清证明。此外,用从 1207 只绵羊收集的血清测定了重组蛋白在 ELISA 中的效率。最后,重组TmHSP70抗原在Western Blot中的敏感性和特异性值为100%,而ELISA的敏感性和特异性分别为66.6%和52.6%。同时,在 1207 份分析的血清样本中,有 567 份(46.97 %)通过 ELISA 检测呈阳性。综上所述,

更新日期:2021-07-22
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