本文關(guān)鍵詞： 手足口病 病原學(xué) EV71 CVA16 分子分型　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：背景與目的:手足口病(hand,foot and mouth disease,HFMD)是由人的腸道病毒感染引起的,近年來發(fā)病率和死亡率位于我國(guó)丙類傳染病之首,少數(shù)患者可能出現(xiàn)無菌性腦膜炎、神經(jīng)源性肺水腫以及伴有心肌炎等癥狀,個(gè)別重癥的患兒隨著病情的惡化,可引起嚴(yán)重并發(fā)癥,若治療不及時(shí)甚至?xí)䦟?dǎo)致死亡。目前報(bào)道的與手足口病相關(guān)的腸道病毒大約有20多種,EV71和CVA16是造成手足口病的主要病原體。隨著基因測(cè)序技術(shù)的飛速發(fā)展以及網(wǎng)絡(luò)數(shù)據(jù)資源的共享,以核苷酸為基礎(chǔ)的分子生物學(xué)研究為病毒進(jìn)化研究提供了方便快捷的方法。本研究對(duì)2015年長(zhǎng)春市手足口病病原學(xué)特征進(jìn)行了分析,闡明2015年長(zhǎng)春市引發(fā)手足口病的主要病原體,并對(duì)其中分離到的EV71和CVA16進(jìn)行分子分型與進(jìn)化研究,進(jìn)一步闡明其遺傳進(jìn)化規(guī)律和特點(diǎn),為長(zhǎng)春市手足口病的預(yù)防和控制工作提供一定的參考依據(jù)。材料與方法:本研究共收集了2015年長(zhǎng)春市手足口病標(biāo)本36份,將標(biāo)本接種到RD細(xì)胞,進(jìn)行病毒的分離和鑒定,對(duì)出現(xiàn)典型細(xì)胞病變效應(yīng)(CPE)的標(biāo)本進(jìn)行病毒RNA的提取,分別采用腸道病毒通用引物、EV71和CVA16特異性引物進(jìn)行鑒定。并對(duì)分離得到的EV71和CVA16,擴(kuò)增其VP1序列,將PCR產(chǎn)物回收、測(cè)序,使用Bioedit與MEGA6.0軟件對(duì)分離獲得的EV71和CVA16的VP1序列進(jìn)行核苷酸和氨基酸同源性比對(duì)分析,構(gòu)建系統(tǒng)進(jìn)化發(fā)育樹。結(jié)果與結(jié)論:2015年長(zhǎng)春市手足口病的發(fā)病高峰期主要集中在4-6月份,占全年收集病例的44.4%,與2015年全國(guó)手足口病發(fā)病高峰期相一致;手足口病患者性別構(gòu)成無顯著性差異;發(fā)病年齡主要集中在2-4歲的兒童,占總發(fā)病數(shù)的75%,其中發(fā)病數(shù)最多的是3歲組,占33.3%。從2015年長(zhǎng)春市36份手足口病標(biāo)本中共分離獲得17株腸道病毒,腸道病毒的陽性分離率為47.2%,陽性標(biāo)本中有5株EV71病毒(占29.4%),6株CVA16病毒(占35.3%),其它腸道病毒為6株(占35.3%)。在2015年長(zhǎng)春市手足口病病原體中除了EV71和CVA16占主要部分,其它腸道病毒在手足口病病原譜中也占據(jù)重要的位置。分離獲得的5株EV71,其VP1序列相互間核苷酸和氨基酸的序列同源性較高,核苷酸之間的同源性為96.2%-97.7%;氨基酸彼此間的同源性為99.2%-99.5%,5株EV71的VP1序列未發(fā)現(xiàn)有明顯差別,5株EV71均為C4a亞型,這與近年來中國(guó)大陸各省份和地區(qū)分離的EV71主要的流行優(yōu)勢(shì)株保持一致。分離獲得的6株CVA16,其VP1序列相互間核苷酸和氨基酸序列的同源性較高,其核苷酸相互間的同源性為97.4%-99.9%;氨基酸之間的同源性為99.3%-99.9%,6株CVA16的VP1序列未發(fā)現(xiàn)有明顯差別,6株CVA16均為B2b亞型,這與近年來中國(guó)大陸各省份和地區(qū)分離的CVA16主要的流行優(yōu)勢(shì)株保持一致,沒有不同抗原的新亞型的出現(xiàn),并且較為穩(wěn)定的核酸序列為疫苗的研制提供了較好的機(jī)會(huì)。
[Abstract]:Background & objective: hand foot and mouth disease (HFMD) is caused by human enterovirus infection. In recent years, morbidity and mortality are the first among Class C infectious diseases in China, a few patients may have aseptic meningitis, neurogenic pulmonary edema and accompanied with myocarditis and other symptoms. Can cause serious complications, if treatment is not timely or even lead to death. There are about 20 kinds of enterovirus associated with hand, foot and mouth disease reported at present. EV71 and CVA16 are the main pathogens causing HFMD. With the rapid development of gene sequencing technology and the sharing of network data resources. The molecular biology based on nucleotides provides a convenient and rapid method for the study of viral evolution. The etiological characteristics of hand-foot-mouth disease in Changchun in 2015 were analyzed in this study. In 2015, the main pathogens of HFMD in Changchun were elucidated, and the molecular typing and evolution of EV71 and CVA16 isolated from them were studied, and the rules and characteristics of genetic evolution were further elucidated. Materials and methods: in 2015, 36 specimens of hand, foot and mouth disease were collected and inoculated into Rd cells in Changchun. The virus was isolated and identified, and the virus RNA was extracted from the specimens with typical cytopathic effect. The common primers of enterovirus were used respectively. EV71 and CVA16 specific primers were used to identify and amplify the VP1 sequence of EV71 and CVA16. The PCR products were recovered and sequenced. Bioedit and MEGA6.0 software were used to analyze the nucleotide and amino acid homology of VP1 sequences of EV71 and CVA16. Results and conclusion: in 2015, the peak period of HFMD in Changchun was mainly in April-June, accounting for 44.4% of the cases collected in the whole year. It was consistent with the peak period of hand, foot and mouth disease in China in 2015. There was no significant difference in gender composition in patients with hand, foot and mouth disease. The age of onset was mainly in children aged 2 to 4 years, accounting for 75% of the total incidence, and the most common incidence was in the group of 3 years old. In 2015, 17 strains of enterovirus were isolated from 36 specimens of hand, foot and mouth disease in Changchun, and the positive isolation rate of enterovirus was 47.2%. Among the positive specimens, there were 5 strains of EV71 virus (29. 4%) and 6 strains of CVA16 virus (35. 3%). In 2015, EV71 and CVA16 were the main pathogens of HFMD in Changchun. Other enterovirus play an important role in the pathogeny of HFMD. The VP1 sequences of 5 strains EV71were highly homologous to each other in nucleotide and amino acid sequences. The homology between nucleotides was 96.2-97.7; The homology of amino acids was 99.2-99.5. The VP1 sequences of 5 EV71 strains showed no significant difference. The 5 strains of EV71 were all C4a subtypes. This was consistent with the main dominant strains of EV71 isolated from provinces and regions in mainland China in recent years. 6 CVA16 strains were isolated. The homology of nucleotide and amino acid sequence between VP1 sequence and amino acid sequence was high, and the homology of nucleotide sequence was 97.4- 99.9. The homology of amino acids was 99.3-99.9. There was no significant difference in the VP1 sequence of 6 CVA16 strains. The six CVA16 strains were all B2b subtypes. This is consistent with the main dominant strains of CVA16 isolated from provinces and regions in mainland China in recent years, and there are no new subtypes of different antigens. And relatively stable nucleic acid sequence provides a good opportunity for vaccine development.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R725.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 杜志成;張王劍;郝元濤;;中國(guó)大陸氣象因素與手足口病發(fā)病相關(guān)關(guān)系的meta分析[J];中國(guó)衛(wèi)生統(tǒng)計(jì);2016年05期

2 楊霞;陳麗紅;單大軍;蘇欣;張?zhí)鞐?鄒杰靜;呂慧敏;;2014—2015年吉林地區(qū)手足口病原學(xué)監(jiān)測(cè)結(jié)果分析[J];中國(guó)衛(wèi)生產(chǎn)業(yè);2016年21期

3 桂瑩;;聯(lián)合干擾素及炎琥寧對(duì)小兒手足口病的治療效果觀察[J];中國(guó)現(xiàn)代藥物應(yīng)用;2016年12期

4 付華;邢劍俠;李艷靜;馬艷玲;溫博;;免疫球蛋白聯(lián)合匹多莫德和利巴韋林治療兒童重癥手足口病的臨床研究[J];現(xiàn)代藥物與臨床;2016年05期

5 朱祺;沈金花;陳文花;顧士康;陸紅梅;;上海松江區(qū)≤6歲兒童家長(zhǎng)對(duì)EV71疫苗接種態(tài)度及影響因素[J];中國(guó)公共衛(wèi)生;2016年07期

6 ;全球首個(gè)腸道病毒71型滅活疫苗正式投入市場(chǎng)[J];臨床合理用藥雜志;2016年08期

7 鄭華珍;劉新光;趙煒;;ELISA檢測(cè)血清EV71-IgM診斷手足口病價(jià)值的Meta分析[J];國(guó)際檢驗(yàn)醫(yī)學(xué)雜志;2016年04期

8 司魯瑩;溫紅玲;王志玉;;腸道病毒71型的分子流行病學(xué)研究進(jìn)展[J];國(guó)際病毒學(xué)雜志;2012年03期

9 李東力;易彬樘;;手足口病流行病學(xué)與防控對(duì)策[J];沈陽部隊(duì)醫(yī)藥;2008年06期

10 ;2008年手足口病預(yù)防控制指南[J];中華實(shí)驗(yàn)和臨床感染病雜志(電子版);2008年03期

相關(guān)博士學(xué)位論文 前1條

1 于品;腸道病毒71型手足口病重癥發(fā)病機(jī)制研究[D];北京協(xié)和醫(yī)學(xué)院;2014年

，

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