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目的对无锡1月份暴发的一起诺如病毒急性胃肠炎疫情进行病原基因测序分析,以鉴定其病原体,并对其基因进行进一步分型和分析。方法采用实时荧光定量RT-PCR法对10份疫情样本进行诺如病毒I型和II型检测,诺如病毒II型阳性样本经半巢氏RT-PCR扩增,然后进行产物纯化、测序,基因数据库比对,并用Clustal X、MEGA 5.1等分析软件构建序列进化树。结果 10份疫情样本中7份样本测出诺如病毒II型阳性,阳性样本经基因扩增、测序,所获序列经基因数据库比对后发现3个为GII.4型,其余为GII.7型,系统进化树分析显示本次病毒株的RdRp区和衣壳区的N/S区的同源分类相同,证明了没有重组株的存在。结论序列测定和分析结果证实这起无锡急性胃肠炎疫情是GII.4型和GII.7型诺如病毒混合感染所致。
Objective To analyze the pathogenic gene sequencing of a Norovirus gastroenteritis outbreak in Wuxi in January in order to identify its pathogens and to further genotype and analyze its gene. Methods Norovirus type I and type II were detected in 10 outbreak samples by real-time fluorescence quantitative RT-PCR. Norovirus type II positive samples were amplified by semi-nested RT-PCR. Then the products were purified and sequenced. Database alignment, and using Clustal X, MEGA 5.1 and other analysis software to build sequence evolution tree. Results Seven out of 10 epidemic samples were positive for Norovirus type II. The positive samples were amplified by gene and sequenced. The obtained sequences were found to be GII.4 type by genetic database and the rest were GII.7 Phylogenetic tree analysis showed that the homology classification of the RdRp region and the N / S region of the capsid region of this strain was the same, which proves that there is no recombinant strain. Conclusion The results of sequencing and analysis confirmed that the epidemic situation of acute gastroenteritis in Wuxi was caused by mixed infection of GII.4 and Norovirus of GII.7 type.