基于基因序列聚类的甲型流行性感冒病毒H3抗原变异规律研究

目的 利用聚类分析方法探讨全球甲型流行性感冒病毒(流感)H3亚型抗原的变异规律。方法下载NCBI GenBank和流感病毒数据库中全部的甲型流感病毒RNA4节段H3亚型基因序列,在Clustalx中进行序列对齐后,使用两阶段聚类法进行分析,并随后探讨各类的三间分布。结果 所有序列可被分为10类,其中7类主要为人流感病毒,人流感病毒和鸟类、其他哺乳动物的流感病毒被明确的分为不同类别,但和猪流感病毒则共存于数个类中。各类呈现出明显的时间分布和宿主分布规律,但并未发现地域分布规律。结论 由于受到人类免疫系统的选择压力,H3抗原呈现出5-7年出现一次较大变异的流行特征,且这一趋势随着近十年来流感疫苗的普遍使用而呈现加速趋势。同时,猪流感病毒和人流感病毒出现在同一类别中,两者的遗传距离较近,这为猪作为病毒重配的载体提供了新的佐证。     

甲型H1N1流行性感冒病毒血凝素蛋白基因进化研究

Objective To determine the evolutionary rate and divergence time of influenza A virus HA gene isolated recently worldwide pandemic and explore the origin and its transmission. Methods A total of 344 HI sequences available in the GenBank (including 248 isolated from human, 84 from swine, 11 from avian, and 1 from ferret) and 7 isolated in Shanghai were collected. The nucleotide substitution rate and time to most recent common ancestor (tMRCA) was calculated using molecular clock theory and Bayesian Skyline Plot (BSP) based on Markov chain Monte Carlo. Then genetic phylogeny was constructed referring to posterior distribution. Results It was found that H1 sequences in the US from human, swine and avian were clustered significantly with swine H1 ones from Asia phylogenetieally (Cluster US). The second cluster (Cluster Eurasian Human) nearly consisted of human H1 sequences isolated in other regions. The third cluster (Cluster Eurasian Animal) consisted of swine and avian H1 sequences from China and Italy respectively. As for all the H1 sequences, the evolutionary rate was of 2.57×10-3substitutions/site per year averagely (95% Highest Posterior Density: 1.96×10-3-3.03×10-3/site per year). The estimated dates for tMRCA of human H1 in Europe and swine H1 in the mainland of China were the earliest, with the corresponding rates of 6.46×10-3/site per year and 0.97×10-3/site per year respectively. The tMRCAs of human and swine H1 sequences from the US were similar, with the rates of 5.86×10-3/site per year and 5.02×10-3/site per year. Conclusion The present flu outbreak was possibly induced by long-term circulation of influenza A virus (H1 N1) in human population and swine herds in America. There was no evidence proving that influenza virus in China involved in the present outbreak.     

上海地区水痘带状疱疹病毒基因型研究

目的：了解上海地区水痘-带状疱疹病毒基因型。方法：用Vero和Vero-E6细胞分离培养临床诊断水痘病人的咽拭子标本,观察病毒的致细胞病变效应（CPE）,对病变细胞培养液提取DNA,进行聚合酶链反应（PCR）,对PCR产物测序,将测序结果与Genbank中公布的水痘病毒序列比对判定水痘带状疱疹病毒和病毒的基因型。结果：PCR产物进行电泳,在550 bp处可见目的片段。测序结果证实为水痘带状疱疹病毒,基因型为J型。结论：从2006年5月华山医院临床诊断水痘患者咽拭子标本分离到的水痘带状疱疹病毒基因型为J型,ORF22读码框与水痘带状疱疹减毒活疫苗V-Oka株基因同源。     

戊型肝炎基因重组研究

目的探讨戊型肝炎病毒基因重组及其对病毒抗原性的影响。方法收集HEV基因全序列,经过对齐、去除空位处理后,运用RDP2.0软件进行筛查,然后利用SimPlot3.5.1软件进行相似性分析和BOOTSCAN分析,对基因重组加以验证,并构建最大似然比进化树,观察重组片断拓扑结构的改变。结果利用RDP2.0软件共检测到3个重组序列,分别是AF459348、D11093、D11092,都是Ⅰ型HEV。相似性分析和BOOTSCAN分析都显示D11093、D11092重组是有统计学意义的,重组片断最大似然进化树的拓扑结构也发生了改变,都发生在Ⅰ型ORF1区。地缘分析表明D11092是基因重组的产物,而D11093极有可能是实验室内发生的重组。结论HEV进化过程中也存在重组,主要发生在Ⅰ型ORF1区;HEV基因重组对于未来HEV抗原性改变可能影响不大。     

甲型H1N1流行性感冒病毒血凝素蛋白基因进化研究

Objective To determine the evolutionary rate and divergence time of influenza A virus HA gene isolated recently worldwide pandemic and explore the origin and its transmission. Methods A total of 344 HI sequences available in the GenBank (including 248 isolated from human, 84 from swine, 11 from avian, and 1 from ferret) and 7 isolated in Shanghai were collected. The nucleotide substitution rate and time to most recent common ancestor (tMRCA) was calculated using molecular clock theory and Bayesian Skyline Plot (BSP) based on Markov chain Monte Carlo. Then genetic phylogeny was constructed referring to posterior distribution. Results It was found that H1 sequences in the US from human, swine and avian were clustered significantly with swine H1 ones from Asia phylogenetieally (Cluster US). The second cluster (Cluster Eurasian Human) nearly consisted of human H1 sequences isolated in other regions. The third cluster (Cluster Eurasian Animal) consisted of swine and avian H1 sequences from China and Italy respectively. As for all the H1 sequences, the evolutionary rate was of 2.57×10-3substitutions/site per year averagely (95% Highest Posterior Density: 1.96×10-3-3.03×10-3/site per year). The estimated dates for tMRCA of human H1 in Europe and swine H1 in the mainland of China were the earliest, with the corresponding rates of 6.46×10-3/site per year and 0.97×10-3/site per year respectively. The tMRCAs of human and swine H1 sequences from the US were similar, with the rates of 5.86×10-3/site per year and 5.02×10-3/site per year. Conclusion The present flu outbreak was possibly induced by long-term circulation of influenza A virus (H1 N1) in human population and swine herds in America. There was no evidence proving that influenza virus in China involved in the present outbreak.     

急性日本血吸虫病与血清白介素?33的关系研究

目的 目的 检测急性日本血吸虫病患者外周血血清白介素?33 （IL?33） 水平, 探讨IL?33在急性日本血吸虫病疾病进展中的作用。方法 方法 选择来自湖区的4例急性日本血吸虫病患者和来自非疫区的15例对照者作为研究对象, 收集人口学资料并采集肘静脉血。采用酶联免疫吸附试验检测所有研究对象的外周血血清IL?33水平。应用Stata 10.0软件进行数据的统计学分析。 结果 结果 急性日本血吸虫病患者血清IL?33水平为517.33 （334.65,1 056.88）pg/ml, 显著高于对照者血清IL?33 水平1.66 （1.66,6.35）pg/ml （Z= -3.207,P = 0.001）。急性日本血吸虫病患者血清IL?33水平与外周血嗜酸粒细胞计数、 患者感染时长的相关系数均为0.8, 相关系数均无统计学意义 （P = 0.2）。结论 结论 IL?33在急性日本血吸虫病患者血清中显著升高, 在日本血吸虫病急性期发挥了促进炎症作用, 并可能在患者感染日本血吸虫后7~9周参与启动Th2型免疫应答。     

70对母婴麻疹野病毒中和抗体检测结果分析

[目的]了解母婴的麻疹野病毒中和抗体水平及其相关性。[方法]采用中和试验的方法直接检测孕妇及新生儿的麻疹野病毒中和抗体水平并进行相关分析。[结果]孕妇及新生儿的麻疹野病毒中和抗体阳性率分别为91．52％和88．57％,平均几何滴度（GMT）分别为61．32和58．17;配对母婴的麻疹野病毒中和抗体水平呈高度正相关（r=0．899,P〈O．01）。当母体的抗体滴度≥l：16时,其子代抗体的阳性率可达100．00％。[结论]母婴的麻疹抗体水平密切相关,提高母源麻疹抗体水平应是控制低月龄小儿麻疹发病的重要措施之一。     

2007-2010年上海市静安区儿童手足口病情况分析和病原学研究

目的 分析上海市静安区2007 -2009年手足口病发病情况,并分析2009-2010年静安区监测点儿童手足口病病原型别及肠道病毒71型(enterovirus 71,EV71) VP1区基因特征.方法 分析上海市静安区2007 -2009年手足口病流行概况;收集2009年4月～2010年12月辖区内儿童医院、幼托机构手足口病患儿咽拭标本,并进行实时荧光RT-PCR检测,确定手足口病病原型别.选择2010年不同月份共19份(含1份重症病例)EV71 PCR阳性标本,接种Vero细胞进行病毒分离培养,并设计特异引物对EV71 VP1全序列进行核苷酸序列测定、比对,构建系统发生树.结果 2008年静安区手足口病发病率明显增高,2009年发病数稳中略降,病例以发生在幼托机构为主.2009年病例咽拭标本实时荧光RT-PCR检测结果显示,柯萨奇病毒A组16型(coxsackievirus A16,CoxA16)阳性率为51.0％ (52/102),EV71阳性率为15.7％ (16/102),其他肠道病毒属阳性率2.9％ (3/102);2010年CoxA16和EV71阳性率均为31.3％ (36/115),其他肠道病毒属阳性率10.4％( 12/115).19株EV71分离株与2008年安徽阜阳EV71分离株同属C基因型中的C4a亚型,核苷酸同源性达95.0％ ～97.1％,氨基酸同源性为95.1％ ～98.3％.结论 静安区手足口病发病率2008年明显增高,2009年发病数稳中略降.2009-2010年静安区手足口病主要病原体为EV71和CoxA16,其他肠道病毒属引起的手足口病有所增加,需加强对其他肠道病毒属的鉴定.     

常用手消毒剂对人肠道病毒71型灭活效果的研究

目的了解常用手消毒剂对人肠道病毒71型(HEV71)的灭活效果。方法采用悬液定量试验法进行试验观察。结果 A类消毒凝胶和B类手部消毒液分别作用1min和5min,对HEV71灭活对数值均>3.0;C类医用复方手消毒剂分别作用1min和5min,对HEV71灭活对数值均>4.0;自制乙醇甘油消毒液作用1min,对悬液内HEV71灭活对数值达5.5。结论目前市场销售常用手消毒剂对人肠道病毒均能达到消毒效果,自制乙醇甘油(77%乙醇、2%甘油)手部消毒液对人肠道病毒的杀灭效果理想。