贵州省25株狂犬病病毒核蛋白基因序列分析

分析贵州省25株狂犬病病毒的核蛋白基因(N基因)序列,探讨贵州省狂犬病流行特征与狂犬病病毒变异情况。以RT-nested PCR检测来自贵州省2005年至2010年不同地区的病人脑组织、病人唾液以及犬脑组织标本狂犬病病毒RNA,经测序与拼接后得到25条N基因全长序列,采用生物信息学软件对N基因序列进行分析。25株狂犬病病毒核蛋白在核苷酸及氨基酸水平上彼此的同源性分别为89.3%～100%和98.%～100%;与国内其他省已发表基因1型狂犬病病毒核苷酸和氨基酸序列同源性分别为88%～99.1%和88%～99.7%,与已知的基因1型狂犬病病毒比较,25株病毒核蛋白氨基酸序列发生了若干位点的取代。进化树分析显示,同一地区内与相邻地区,以及同一时间段与相邻时间段内狂犬病病毒N基因进化亲缘关系相近。25株贵州省狂犬病病毒流行毒株均属基因1型,其核蛋白在基因的核苷酸及推导的氨基酸水平上均有变异,且这些变异具有地域和时间分布特性。

Analysis on nucleoprotein gene sequence of 25 rabies virus isolates in Guizhou Province, China

To analyze 25 nucleoprotein gene (N gene) sequences of rabies viruses circulating in Guizhou province during 2005-2010, China, and to explore the epidemic characteristics and the probable mutant of rabies in Guizhou Province. Rabies virus RNA in human brain tissues, human saliva, and domestic dog brain tissues derived from different prefectures of Guizhou Province were detected with RT-nested PCR, and the amplified products were then sequenced. Bioinformatics software was used to determine the genetic characteristics of these rabies viruses. The sequences of N gene of 25 Guizhou provincial isolates were identical with homogeny between 97.5% - 99.3% and 98.4% - 99.8% at nucleotide and deduced amino acid level, respectively, while the identities between them and isolated strains from other province of China were 88% - 99.1% and 88% - 99.7%. There were several amino acid substitutions in the nucleoprotein of 25 Guizhou isolates compared with the known genotype 1 isolates. The analysis of phylogenetic tree of 25 Guizhou isolates was demonstrated to be genetically divided into two groups, indicating that the virus presented a unique characteristics in geographic distribution and in a time dependent-manner. And phylogenetic tree of 25 Guizhou isolates and 7 genotype 1 strains isolated from other Province of China was also divided into two groups, which were further composed of several subgroups, respectively. From these observations, the rabies viruses derived from Guizhou province were still genotype 1. These isolates of rabies virus were diverged from the strains isolated from other provinces in both gene sequences and deduced amino acid sequences, and these divergences were characterized in geographic distribution and in a time-dependent manner.