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目的了解河南省流行的麻疹野病毒的基因特征和型别,为制定消除麻疹策略提供依据。方法用非洲绿猴肾细胞/淋巴信号激活因子转染的非洲绿猴肾细胞(Vero/SLAM)从麻疹疑似病例标本中分离麻疹野病毒,用逆转录-聚合酶链反应(RT-PCR)扩增麻疹野病毒分离株的核蛋白(N)基因羧基末端676个核苷酸片段,再对扩增产物进行核苷酸序列测定和分析。以N基因羧基末端450个核苷酸片段构建基因系统进化树,进行遗传距离研究;根据核苷酸序列推导出氨基酸序列,分析氨基酸序列变异情况。结果对2010-2011年分离的49株麻疹病毒进行测序后显示均为H1a基因型,有较高的同源性,大部分序列出现4个核苷酸变异,出现位置在139(G→A)、244(A→G)、298(C→A)、365(G→A)位;推导出的氨基酸序列与H1a基因型比较发现,编码氨基酸422(G→S)、457(S→G)、475(L→I)、497(R→K)位点的变异。结论河南省2010-2011年麻疹野病毒流行优势株为H1a基因型,但存在一定的进化距离,部分分离株氨基酸序列发生突变,呈现多个传播链的流行。
Objective To understand the genetic characteristics and genotypes of wild measles virus in Henan Province, and to provide basis for formulating measles elimination strategy. Methods The wild type of measles virus was isolated from the suspected cases of measles virus using African green monkey kidney cells (Vero / SLAM) transfected with African green monkey kidney cell / lymph node activator and reverse transcriptase-polymerase chain reaction (RT-PCR) The 676 nucleotides of the carboxyl terminal of the nucleoprotein (N) gene of the wild measles virus were amplified, and the amplified products were sequenced and analyzed. The 450 nucleotide sequences of the N gene were used to construct the phylogenetic tree to study the genetic distance. The amino acid sequence was deduced based on the nucleotide sequence and the amino acid sequence variation was analyzed. The results showed that all of the 49 measles viruses isolated from 2010 to 2011 showed H1a genotypes with high homology, most of them showed 4 nucleotide variations and appeared at 139 (G → A) (G → S), 457 (S → G), 244 (A → G), 298 (C → A) and 365 (G → A) , 475 (L → I), 497 (R → K) loci. Conclusion The epidemic predominant strain of wild measles virus from 2010 to 2011 in Henan Province is H1a genotype, but there is a certain evolutionary distance. Some amino acid sequences of some isolates mutate and show the spread of multiple transmission chains.