山西省2004～2005年流感病毒的流行株基因变异及抗体水平研究

发布时间：2018-04-23 12:08

本文选题：流感病毒 + 核苷酸序列测定　；参考：《山西医科大学》2006年硕士论文

【摘要】： 目的:流行性感冒(influenza)是由流感病毒引起的一种常见的急性呼吸道传染病,流感难以控制的主要原因是病毒抗原性尤其是血凝素极易发生变异,使人群通过感染或疫苗获得的免疫力对新流行株失去作用。本研究对山西省2004～2005年流行期流感的流行进行了病原学、血清学等方面的研究。为进一步研究山西省流感病毒的型别变迁规律,及时抓住有意义的变异株,指导今后流感的监测、诊断及疫苗研制,搞好流感疫情的预测预报和及时采取有效防制措施提供理论依据。方法2004年10月至2005年3月,将山西省人民医院,山西医科大学第一附属医院,山西省儿童医院内科和儿科门诊中的流感样病例做为研究对象,收集其基本人口学特征资料。并且对发病三天以内,没有服用过抗病毒药物的193例流感样病例,采集其口咽部位拭子标本,接种于狗肾细胞(MDCK)分离流感病毒,通过微量血凝抑制(HI)实验进行型别鉴定,用RT-PCR扩增其HA1区基因,对扩增产物进行序列测定,与Genebank中的10株代表株进行序列分析及进化分析。同时于2005年3月份流感流行后期,在山西省从北向南依次抽取大同、晋中、临汾三地区,在这三个地区的市级医院门诊病人中分别选择了133、128、132名未接受过流感疫苗接种的非呼吸道疾病的病人,采集其空腹静脉晨血,通过微量血凝抑制(HI)实验对其体内流感病毒抗体进行检测分析。用SPSS11.0软件对资料进行χ2检验,Kruskal-Wallis H秩和检验等。HA1区基因序列分析等采用DNASTAR5.0与GENEDOC软件。结果(1)2004～2005年流感流行期,流感样病例数占门诊总病例数的0.23%,就诊高峰在2月份。(2)193例疑似流感样患者中共分离出13株B型流感病毒,1月份与2月份病毒分离率最高。(3)HA1基因核苷酸及其推导的氨基酸序列及基因进化分析表明,13株流感病毒均为Yamagata样流感病毒,属于B/Shanghai/361/2002(2001～2004年全国流行株代表株,WHO推荐的该年度疫苗组分)分化出的种系,但与B/Jiangsu/10/03同源性最高。(4)13株毒株与B/Shanghai/361/2002相比在抗原结合B、D位点及其附近发生了变异,糖基化位点的数目也发生了改变。(5)山西省人群血清抗体B型抗体总阳性率为95.9%、几何平均滴度(GMT)为190.27,均高于A3( 91.6%,46.27)、A1 (85.8%,35.58)两型,三型抗体的阳性率与GMT的分布在不同年龄与地区间有差异。结论(1)山西省2004～2005年流感的发生与全国大的流行趋势基本一致, B型流感病毒成为流行优势株。(2)所分离的B型流感病毒与B/Shanghai/361/2002相比基因发生了变异,但未达到抗原漂移。(3)经过2004～2005年流感流行山西省人群对B型流感病毒已形成了一定的抵抗力,对A型流感病毒的抵抗力较低,应警惕B型流感病毒进一步发生变异以及A型流感病毒引起流行甚至爆发的可能。
[Abstract]:Objective: influenza influenzas is a common acute respiratory infection caused by influenza virus. The main reason of influenza is that the antigenicity of influenza virus, especially hemagglutinin, is easy to mutate. The immunity acquired by an infection or vaccine in a population is rendered ineffectual to a new epidemic strain. In this study, the epidemiology and serology of influenza in the epidemic period from 2004 to 2005 in Shanxi Province were studied. In order to further study the pattern changes of influenza viruses in Shanxi Province, seize the meaningful variant strains in time and guide the surveillance, diagnosis and vaccine development of influenza in the future, To do well the forecast of influenza epidemic situation and to take effective prevention and control measures in time to provide the theoretical basis. Methods from October 2004 to March 2005, the influenza-like cases from the people's Hospital of Shanxi Province, the first affiliated Hospital of Shanxi Medical University, the Department of Internal Medicine and the Pediatric outpatient Department of Shanxi Children's Hospital were selected as the research objects, and their basic demographic characteristics were collected. The influenza virus was isolated from the oral and pharyngeal swabs of 193 influenza-like patients who had not taken antiviral drugs within three days. The influenza virus was isolated by inoculation with MDCK in dog kidney cells, and the type of influenza virus was identified by microhemagglutination inhibition test. The HA1 region gene was amplified by RT-PCR. The amplified products were sequenced and sequenced with 10 representative strains in Genebank. At the same time, in the late period of influenza epidemic in March 2005, Datong, Jinzhong and Linfen were drawn from north to south in Shanxi Province. Among the outpatients in municipal hospitals in these three regions, 133128132 patients with non-respiratory diseases who had not been vaccinated against influenza were selected, and their fasting morning blood was collected. The antibody to influenza virus in vivo was detected and analyzed by microhemagglutination inhibition (HI) test. DNASTAR5.0 and GENEDOC software were used to analyze the gene sequence of Kruskal-Wallis H region by using SPSS11.0 software. Results the influenza epidemic period from 2004 to 2005, Influenza-like cases accounted for 0.23% of the total outpatient cases. The peak was in February. 2193 suspected influenza-like patients isolated 13 strains of influenza B virus in January and February with the highest isolation rate of HA1 gene nucleotide and its derivation. Amino acid sequence and gene evolution analysis showed that all of the 13 influenza viruses were Yamagata like influenza viruses. It belongs to the strain of B / Shanghai / 361R / 2002 / 2002 / 20012004, which is recommended by WHO as the vaccine component of this year, but has the highest homology with B/Jiangsu/10/03. Compared with B/Shanghai/361/2002, the strain of 13 strains has mutated at and around the antigen-binding B/Jiangsu/10/03 D site. The number of glycosylation sites also changed.) the total positive rate of serum antibody type B antibody in Shanxi population was 95.9 and the geometric mean titer of GMTwas 190.27, which was higher than that of A3 (91.6% 46.27%) and 85.8% (35.58%). The positive rate of type III antibody and the distribution of GMT were different in different ages and regions. Conclusion (1) the occurrence of influenza in Shanxi Province from 2004 to 2005 is basically consistent with the trend of epidemic in China. Influenza B virus has become the dominant epidemic strain, and the gene of influenza B virus isolated from Shanxi Province is different from that of B/Shanghai/361/2002. However, after 2004 ~ 2005 influenza epidemic population in Shanxi Province has developed a certain resistance to influenza B virus, but the resistance to influenza A virus is relatively low. We should be on guard against the further variation of influenza B virus and the possibility of epidemic and even outbreak caused by influenza A virus.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R181.3;R373

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