Citation: TANG Jia-Qi, CAO Min, WANG Chang-Jun, LEI Mo-Li, WEI Chun-Bao, XIE Chun-Yan. Studies on Preparations of Nucleocapsid Protein Recombinant Antigen and Genotyping of Hantaviruses .VIROLOGICA SINICA, 1999, 14(4) : 314-321.

Studies on Preparations of Nucleocapsid Protein Recombinant Antigen and Genotyping of Hantaviruses

  • Available online: 05 December 1999
  • Two groups of primers were designed according to the gene backgrounds of prototype virus of HTN and SEO serotypes and corrected by co mputer.One group of primers was used to elone entire S gencane segment and partial S genome segment with respect to N.terrmnal,n 1 two cloned genes were fusionally expressed and non-fusionally expressed by 27 system .The non fusionally expressed products whose working co ncentration were 1:10 000 presented a good bi0_ logical activity though their yields were lower than the fusionally expressed products.The other group of primers was used to establish a meth0d of RTPCR to detect RNAs of 37 virus isolates of HFRSv 2 positeve standard viruses and 5 negtive controls. On comparision with that of cELISA,the detecting rates of two m ethods were 100% and 84.6% respectively,the co incidental ratewas 84.6% whilethefoiT~1er had 15.4% higher sensitivitythan thelatter.Thetyping method of RFLP was set up by digesting the PCR products of 20 virus isolates with Ras I and HindUl resulting that 9 OUt of the tot81 were HTN.8 were SEO aM 3 were n0t determined re spectively.The same 20 viruses have been previoushy typed using serotyping method resulting tha t only 11 could he typed successfully.showing a high co nsistency with that of RTPCR-RFLP method and a 30% lower typing etficiency than the latter.

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    Studies on Preparations of Nucleocapsid Protein Recombinant Antigen and Genotyping of Hantaviruses

    • 1. MillmryReserch Institute,0rMedicine and Technology of NanjingCommand ,Nanling 210002

    Abstract: Two groups of primers were designed according to the gene backgrounds of prototype virus of HTN and SEO serotypes and corrected by co mputer.One group of primers was used to elone entire S gencane segment and partial S genome segment with respect to N.terrmnal,n 1 two cloned genes were fusionally expressed and non-fusionally expressed by 27 system .The non fusionally expressed products whose working co ncentration were 1:10 000 presented a good bi0_ logical activity though their yields were lower than the fusionally expressed products.The other group of primers was used to establish a meth0d of RTPCR to detect RNAs of 37 virus isolates of HFRSv 2 positeve standard viruses and 5 negtive controls. On comparision with that of cELISA,the detecting rates of two m ethods were 100% and 84.6% respectively,the co incidental ratewas 84.6% whilethefoiT~1er had 15.4% higher sensitivitythan thelatter.Thetyping method of RFLP was set up by digesting the PCR products of 20 virus isolates with Ras I and HindUl resulting that 9 OUt of the tot81 were HTN.8 were SEO aM 3 were n0t determined re spectively.The same 20 viruses have been previoushy typed using serotyping method resulting tha t only 11 could he typed successfully.showing a high co nsistency with that of RTPCR-RFLP method and a 30% lower typing etficiency than the latter.

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