本文選題：流感病毒 + 核苷酸序列測(cè)定��； 參考：《山西醫(yī)科大學(xué)》2006年碩士論文

【摘要】： 目的:流行性感冒(influenza)是由流感病毒引起的一種常見的急性呼吸道傳染病,流感難以控制的主要原因是病毒抗原性尤其是血凝素極易發(fā)生變異,使人群通過感染或疫苗獲得的免疫力對(duì)新流行株失去作用。本研究對(duì)山西省2004～2005年流行期流感的流行進(jìn)行了病原學(xué)、血清學(xué)等方面的研究。為進(jìn)一步研究山西省流感病毒的型別變遷規(guī)律,及時(shí)抓住有意義的變異株,指導(dǎo)今后流感的監(jiān)測(cè)、診斷及疫苗研制,搞好流感疫情的預(yù)測(cè)預(yù)報(bào)和及時(shí)采取有效防制措施提供理論依據(jù)。 方法2004年10月至2005年3月,將山西省人民醫(yī)院,山西醫(yī)科大學(xué)第一附屬醫(yī)院,山西省兒童醫(yī)院內(nèi)科和兒科門診中的流感樣病例做為研究對(duì)象,收集其基本人口學(xué)特征資料。并且對(duì)發(fā)病三天以內(nèi),沒有服用過抗病毒藥物的193例流感樣病例,采集其口咽部位拭子標(biāo)本,接種于狗腎細(xì)胞(MDCK)分離流感病毒,通過微量血凝抑制(HI)實(shí)驗(yàn)進(jìn)行型別鑒定,用RT-PCR擴(kuò)增其HA1區(qū)基因,對(duì)擴(kuò)增產(chǎn)物進(jìn)行序列測(cè)定,與Genebank中的10株代表株進(jìn)行序列分析及進(jìn)化分析。同時(shí)于2005年3月份流感流行后期,在山西省從北向南依次抽取大同、晉中、臨汾三地區(qū),在這三個(gè)地區(qū)的市級(jí)醫(yī)院門診病人中分別選擇了133、128、132名未接受過流感疫苗接種的非呼吸道疾病的病人,采集其空腹靜脈晨血,通過微量血凝抑制(HI)實(shí)驗(yàn)對(duì)其體內(nèi)流感病毒抗體進(jìn)行檢測(cè)分析。用SPSS11.0軟件對(duì)資料進(jìn)行χ2檢驗(yàn),Kruskal-Wallis H秩和檢驗(yàn)等。HA1區(qū)基因序列分析等采用DNASTAR5.0與GENEDOC軟件。 結(jié)果(1)2004～2005年流感流行期,流感樣病例數(shù)占門診總病例數(shù)的0.23%,就診高峰在2月份。(2)193例疑似流感樣患者中共分離出13株B型流感病毒,1月份與2月份病毒分離率最高。(3)HA1基因核苷酸及其推導(dǎo)的氨基酸序列及基因進(jìn)化分析表明,13株流感病毒均為Yamagata樣流感病毒,屬于B/Shanghai/361/2002(2001～2004年全國(guó)流行株代表株,WHO推薦的該年度疫苗組分)分化出的種系,但與B/Jiangsu/10/03同源性最高。(4)13株毒株與B/Shanghai/361/2002相比在抗原結(jié)合B、D位點(diǎn)及其附近發(fā)生了變異,糖基化位點(diǎn)的數(shù)目也發(fā)生了改變。(5)山西省人群血清抗體B型抗體總陽(yáng)性率為95.9%、幾何平均滴度(GMT)為190.27,均高于A3( 91.6%,46.27)、A1 (85.8%,35.58)兩型,三型抗體的陽(yáng)性率與GMT的分布在不同年齡與地區(qū)間有差異。 結(jié)論(1)山西省2004～2005年流感的發(fā)生與全國(guó)大的流行趨勢(shì)基本一致, B型流感病毒成為流行優(yōu)勢(shì)株。(2)所分離的B型流感病毒與B/Shanghai/361/2002相比基因發(fā)生了變異,但未達(dá)到抗原漂移。(3)經(jīng)過2004～2005年流感流行山西省人群對(duì)B型流感病毒已形成了一定的抵抗力,對(duì)A型流感病毒的抵抗力較低,應(yīng)警惕B型流感病毒進(jìn)一步發(fā)生變異以及A型流感病毒引起流行甚至爆發(fā)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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R181.3;R373

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