Full sequence analysis and characterization of the Shenzhen Norovirus strain SZ2010422 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To obtain information on viral molecular structural and evolutionary characteristics, we conducted the SZ2010422 full-length genomic analysis.</p><p><b>METHODS</b>Primers were designed by New Orleans full sequence, SZ2010422 full genome was amplified by RT-PCR, the whole genome sequence and the capsid domain amino acid sites was analysised after cloned and sequenced.</p><p><b>RESULTS</b>The genome of G II-4 Norovirus SZ2010422 strain was consist of 7559 bp, it revealed three ORFs composites of the whole genome, ORF1 (5100 bp), ORF2 (1623 bp), ORF3 (807 bp) respectively, ORF1 and ORF2 had 19 nucleotide overlap. By evolutionary comparative analysis found SZ2010422 genomic nucleotide sequences with reference strains of G II-4 New Orleans1805 strains the highest homology with a total length of homology was 99.3%, of ORF1 (99.5%), ORF2 (99.2%), ORF3 (98.6%). Phylogenetic analyses showed SZ2010422 belonging to G II-4 New Orleans variant. Date of 541 amino acid analyses showed: New Orleans variant strains of popular sites: aa310N or K, --> S aa341D --> of N, aa359T--> S, aa396H --> P, aa460H --> Y.</p><p><b>CONCLUSION</b>Norovirus SZ2010422 belonged to the G II-4 New Orleans variant. In This study, SZ2010422 full sequence can be used not only as a full-length NoV variant sequence standard for future comparison studies, but also as useful material for the public health field by enabling the diagnosis, vaccine development, and prediction of new emerging variants. Noroviruses; Genes; Sequence analysis</p>

Genetic characterization of G II.12 norovirus in Beijing from 2008 to 2009 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To reveal the genetic characteristics of GII.12 Norovirus strains isolating from stool samples of adults with diarrhea in Beijing during 2008-2009.</p><p><b>METHODS</b>RdRp, ORF2, ORF3 and ORF1/ORF2 overlap region were respectively amplified by primers using RT-PCR. The products were purified, cloned, sequenced and then aligned, phylogenetic and recombinant analyzed by softwares of DNAStar, MEGA and SimPlot.</p><p><b>RESULTS</b>According to the phylogenetic analysis, 11 strains belonged to G II.g in the RdRp region,while GII.12 in the ORF2 and ORF3. SimPlot analysis further confirmed the 11 strains were recombinant strains ( G II.g [RdRp]/G II.12 [capsid]).</p><p><b>CONCLUSION</b>G II.12 Norovirus prevailing in Beijing and other regions of the world belonged to the same strain, and we identified the genetic characteristics of G II.12 Norovirus in Beijing.</p>

Optimizing expression of the capsid protein VP2 from human Bocavirus and establish it's seroepidemiology assying methord / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To obtain sufficient recombinant VP2 protein of human Bocavirus and establish it's seroepidemiology assying metbord. METHORD: Tbe capsid protein VP2 DNA genes of HBoV1 and 2 were optimized in accordance with tbe usage of the favorite codons in K coil so as to enhance its protein expression in prokaryotic expressing system. The protein was purified by Ni-NTA column, and its antigenicity was determined by Western Blot. Then establish ELISA to detect the specific anti-VP2 IgG antibodies against HBoV1 and 2 in healthy children aged 3-6 years in Nanjing, China.</p><p><b>RESULTS</b>The recombinant protein 6 x His-VP2 was produced in a larger quantity at 25 degrees C induced by IPTG (1 mmol/L) over night and purified by Ni-NTA column. Seropositive rates of HBoV1 and 2 were 62.2% and 55.5% and their mixed seropositivity was 37%.</p><p><b>CONCLUSION</b>The optimizing expression of the capsid protein VP2 from human Bocavirus constructed successfully and get a high yield under certain conditions. The established ELISA could be used to further analyze seroepidemiology of HBoV in China.</p>