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Development of an Indirect ELISA for Detection of T.solium Based on Recombinant 18 kDa Protein

Development of an Indirect ELISA for Detection of T.solium Based on Recombinant 18 kDa Protein
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摘要 The gene encoding the 18 kDa protein of Taenia solium metacestodes was amplified by RT-PCR and cloned into the pGEM-T vector for sequencing.The recombinant plasmid named pGEX-CE18 was constructed and transformed into E.coli BL21 for in vitro expression.SDS-PAGE and Western blot were employed for analyzing the recombinant protein,which was then used for development of an indirect ELISA for detection of anti-cysticercosis antibodies.The results showed that the recombinant protein of interest was 35 kDa in size,accounting for 28%of total bacteria proteins,and reacted with positive sera against cysticercosis.Using the newly-constructed indirect ELISA and a commercially available ELISA kit,paired analyses of 178 serum samples indicated that the concordant rate was 98.83%and the ELISA exhibited good specificity and sensitivity,supporting its utility and application for diagnosis of cysticercosis. The gene encoding the 18 kDa protein of Taenia solium metacestodes was amplified by RT-PCR and cloned into the pGEM-T vector for sequencing.The recombinant plasmid named pGEX-CE18 was constructed and transformed into E.coli BL21 for in vitro expression.SDS-PAGE and Western blot were employed for analyzing the recombinant protein,which was then used for development of an indirect ELISA for detection of anti-cysticercosis antibodies.The results showed that the recombinant protein of interest was 35 kDa in size,accounting for 28%of total bacteria proteins,and reacted with positive sera against cysticercosis.Using the newly-constructed indirect ELISA and a commercially available ELISA kit,paired analyses of 178 serum samples indicated that the concordant rate was 98.83%and the ELISA exhibited good specificity and sensitivity,supporting its utility and application for diagnosis of cysticercosis.
出处 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期87-92,共6页 ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金 supported by National High-tech Research and Development Plan(863 Project)(2006AA10A207)
关键词 CYSTICERCUS cellulosae 18 kDa PROTEIN E.COLI EXPRESSION RECOMBINANT PROTEIN ELISA Cysticercus cellulosae 18 kDa protein E.coli expression recombinant protein ELISA
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  • 1殷竹君,张耀娟,章子豪,刘静,史志明,侯敏.抗囊虫单抗的制备及其应用研究[J].中国寄生虫病防治杂志,2003,16(2):77-80. 被引量:11
  • 2da SILVA M R,MAIA A A,ESPINDOLA N M,et al.Recombinant expression of Taenia solium TS14 antigen and its utilization for immunodiagnosis of neurocysticercosis. Acta Tropica . 2006
  • 3ASSANA E,KANOBANA K,TUME C B,et al.Isolation of a 14 kDa antigen from Taenia solium cyst fluid by HPLC and its evaluation in enzyme linked immunosorbent assay for diagnosis of porcine cysticercosis. Research in Veterinary Science . 2007
  • 4GU T C,CHEN Z L.Introduction of Modern Biotechnology. . 2000
  • 5YAO X H,,YANG S M,ZHAO H Y,et al.Porcine cysticercosisspecific antigen extraction and application. Chinese Journal of Veterinary Science and Technology . 1989
  • 6LEE E G,LEE M Y,CHUNG J Y,et al.Feasibility of baculovirusexpressed recombinant 10-kDa antigen in the serodiagnosis of Taenia solium neurocysticercosis. Transactions of the Royal Society of Tropical Medicine and Hygiene . 2005
  • 7ZHAO X H,ZHNEG Y W,ZHU Y N,et al.Detection of serum of patients cerebral cysticercosis clinical of circulating antigen using monoclonal antibody ELISA. Chinese Journal of Parasitology and Parasitic Diseases . 1997
  • 8Hancock K,Pattabhi S,Whitfield FW,Yushak ML.Characterization and cloning of T24, a Taenia solium antigen diagnostic for cysticercosis. Molecular and Biochemical Parasitology . 2006
  • 9JIN D Y,LI M F.Molecular Cloning:A LaboratoryManual. . 1999
  • 10Yoshii,H,Furuta,T,Yonehara,T.Refolding of denatured/reduced lysozyme at high concentration with diafiltration. Bioscience Biotechnology and Biochemistry . 2000

二级参考文献4

  • 1Otero R, Gutierrez JM, Rojas G,et al. A randomized blinded clinical trial of two antivenoms,prepared by caprylic acid or ammonium sulphate fractionation of IgG, in Bothrops and Porthidium snake bites in Colombia :correlation bet ween safety and biochemical characteristics of antivenoms[J]. Toxicon, 1999,37(6) :895--908.
  • 2Tsang VC,Greene RM,Pilcher JB. Optimization of t he covalent conjugating procedure (NaIO4) of horseradish peroxidase to antibodies for use in enzyme -linked immunosorbent assay[J]. J Immunoassay,1995,16(4) :395--418.
  • 3许隆祺,蒋则孝,周长海,张雪强,任正轩,常江,许景田,李远璧,张莉莉,余森海,张广源,马成骥.我国囊尾蚴病分布情况调查[J].中国寄生虫病防治杂志,1999,12(1):30-32. 被引量:34
  • 4王凯慧,徐之杰,甄殿魁.应用免疫印迹技术和ELISA对猪囊尾蚴抗原对比分析[J].中国寄生虫学与寄生虫病杂志,1993,11(1):53-56. 被引量:13

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