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Virologica Sinica, 14 (4) : 314-321, 1999
Studies on Preparations of Nucleocapsid Protein Recombinant Antigen and Genotyping of Hantaviruses
MillmryReserch Institute,0rMedicine and Technology of NanjingCommand ,Nanling 210002
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 2"7 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 RT—PCR 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 RT—PCR-RFLP
method and a 30% lower typing etficiency than the latter.
  Published online: 5 Dec 1999
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