静安区手足口病流行趋势及病原学研究

发布时间：2018-03-23 16:24

本文选题：手足口病　切入点：流行趋势　出处：《复旦大学》2012年硕士论文

【摘要】：本次课题通过收集2005年-2010年静安区手足口病常规报表、监测资料等分析手足口病的流行趋势。应用RT-PCR、细胞培养等实验技术对2009年-2010年监测点手足口病病例标本进行检测,分析引起手足口病的病毒病原型别分布和肠道病毒EV71的VP1基因特征,以了解静安区手足口病的流行规律和分子流行病学特征。通过实验研究不同浓度含氯消毒剂对肠道病毒71型的灭活效果,从而为指导本区手足口病防控措施的制定提供依据。本课题研究内容包括以下几个内容：1.上海市静安区2005-2010年手足口病流行特征分析 2005年发病率为20.44/10万,2006年为18.72/10万,2006年较2005年有所下降。2007年发病率为31.32/10万,2008年为24.84/10万,较2007年有所下降。2009、2010年手足口病的发病率均较2008年有所上升,但2005-2010年静安区本市人口手足口病年发病率以2007年最高。发病高峰集中在每年的5-7月份,2009年11月还出现第二个发病高峰。手足口病的年发病率以位于静安区西北部的曹家渡和江宁街道较高,其他街道总体发病率较低。男女性别比约为1.4：1,5岁以下儿童比例达到83.80%。以幼托机构儿童比例最多。2.上海市静安区2009-2011年手足口病病原监测结果分析 2009-2011年分别收集监测点上海市儿童医院临床诊断手足口病例咽拭、肛拭标本90份、180份和242份,采用Real-time RT-PCR方法进行检测。结果显示,2009-2011年间,监测点儿童手足口病例中柯萨奇病毒A16型(CA16)阳性率逐年下降,而肠道病毒71型(EV71)和肠道病毒属其他病毒(EV)阳性率呈上升趋势。对2010年7月至2011年12月监测点临床诊断手足口病例175例分别采集咽拭和肛拭标本,分别用Real-time RT-PCR方法进行EV71、CA16、EV检测。用McNemar检验比较两种标本检测结果的差异,EV71和EV在咽拭子和肛拭子两种标本检测结果有差异,P值均0.05,CA16在两种标本中的检测结果无显著差异。应用Kappa检验,Kappa值均0.4,显示咽拭子和肛拭子的检测结果基本一致。根据本次实验配对标本检测EV71、CA16、EV结果不一致与标本采集时间进行分析,推断肠道病毒可能在发病2天内在咽部大量繁殖扩增,此时通过呼吸道传播的可能性较大。将2010年7月至2011年12月监测点临床诊断手足口病例采集的175对咽拭和肛拭标本Real-time RT-PCR检测结果按照标本采集时间进行分析,对于EV71来说,咽拭标本在发病后3天、4-11天均可检出阳性,而CA16、EV在发病3天后都无法检出阳性,说明EV71在呼吸道带毒持续时间长于CA16和肠道病毒属病毒。 3.上海市静安区人肠道病毒71型基因进化研究随机选择2010年不同月份19份PCR阳性的标本进行Vero细胞培养,其中1例为重症培养物、18份为非重症培养物,对EV71病毒PCR扩增产物序列(VPl基因1200bp)系统发生分析,所有19株EV71病毒与2007年山东及2008年安徽阜阳EV71分离株同属C基因型中的C4a亚型。将本次研究的19株EV71分离株的VPl基因与从GenBank中检索到EV71各基因型和亚型的代表株序列,包括A、B、Cl、C2、C3、C4和C5亚型进行氨基酸和核苷酸同源性分析,其中C4亚型选择的是2008年安徽省阜阳市流行株。分析发现,19株EV71分离株的VP1基因与C基因型代表株核苷酸同源性在84.3%-97.1%,氨基酸同源性在88.6%-98.3%；与C4基因亚型最为相近,其核苷酸同源性在95.0%-97.1%,氨基酸同源性在95.1%-98.3%。我国自1998年从广东省分离鉴定出C4b亚型EV71分离株后,至2003年国内EV71亚型一直为C4b。2003-2004年,EV71的C4a和C4b两个亚型在我国共存,如浙江、广东、台湾。从2005年至今,上海、河南、云南、北京、山东、安徽、浙江等地分离到的EV71均为C4a亚型,重症和散发病例基因型相同无差异。而美国、澳大利亚、马来西亚、新加坡、日本等国在不同时间段内都曾出现过不同的EV71基因亚型。 4.含氯消毒剂对肠道病毒71型灭活效果的研究肠道病毒71型主要通过粪口途径和密接接触传播。目前,上海地区幼托机构通常使用有效氯浓度为500mg/L的含氯消毒剂对儿童的玩具、毛巾及课桌、门把手等物体表面进行消毒。因此,我们通过试验研究观察了该浓度含氯消毒剂对悬液内EV71病毒的杀灭效果。由于消毒剂对细胞的毒性,本次实验发现有效氯浓度为500mg/L的含氯消毒剂作用5min即可对悬液中的EV71病毒达到至少99.99%的杀灭率；与有效氯浓度为1000mg/L的含氯消毒剂杀灭率相一致。由于试验采用的为病毒悬液,消毒剂与肠道病毒能够直接接触作用,但在日常消毒工作中,含氯消毒剂对包裹在毛巾、衣物中的病毒的杀灭作用会受到物品中有机物、静电吸附等因素的影响。因此,对于怀疑有手足口病患儿接触污染的物品,可通过提高有效氯浓度(1000mg/L或更高)或延长消毒剂作用时间进行消毒处理,保证病毒灭活效果,预防手足口病的传播。
[Abstract]:This topic through the collection of 2005 -2010 Jingan District HFMD routine report, epidemic trend analysis of monitoring data of HFMD. The application of RT-PCR, cell culture techniques were on hand foot mouth disease in 2009 -2010 monitoring points were detected, analyzed VP1 gene distribution type virus and enterovirus EV71 HFMD the cause, in order to understand the epidemiology and molecular epidemiology of HFMD in Jingan District. Through the experimental study of different concentrations of chlorine disinfectant on enterovirus 71 inactivation effect, so as to guide the prevention and control measures in the area of foot and mouth disease to provide basis for the establishment of the research content of the thesis includes the following contents: analysis of Shanghai 1. Jingan District 2005-2010 HFMD epidemic characteristics
In 2005, the incidence rate of 20.44/10 million, 18.72/10 million in 2006 2006, declined in 2005.2007 years the incidence rate of 31.32/10 million, 24.84/10 million in 2008, declined in 2007.20092010 years of HFMD incidence increased compared to 2008, but 2005-2010 years of hand foot mouth disease in Jingan District population of the city the annual incidence rate was highest in 2007. The peak incidence occurred in every 5-7 months, in November 2009 second. The peak incidence of HFMD incidence in the northwest of Jingan District and Jiangning Caojiadu street high street the other overall incidence is relatively low. The proportion of male and female sex ratio for children under the age of 1.4:1,5 years old 83.80%. in preschools of the proportion of children the most.2. of Shanghai city Jingan District 2009-2011 years HFMD pathogen surveillance results
2009-2011 years of monitoring points were collected in Shanghai Children's Hospital clinical diagnosis of HFMD cases of pharyngeal swab, anal swab specimens of 90 copies, 180 copies and 242 copies, using Real-time RT-PCR method. Results showed that 2009-2011 years of monitoring points children HFMD cases in Coxsackie virus type A16 (CA16) positive rate decreased year by year, and enterovirus 71 type (EV71) and other enterovirus virus (EV) positive rate increased.
On July 2010 to December 2011 monitoring of clinical diagnosis of 175 cases of HFMD cases were collected from throat swabs and anal swabs, respectively using Real-time RT-PCR method EV71, CA16, EV detection. Differences with McNemar test comparison of two samples of test results, EV71 and EV in throat swabs and anal swabs of two specimen test results the difference, P value was 0.05, CA16 was detected in two samples showed no significant difference. The application of Kappa test, Kappa value was 0.4, showed that the detection of throat swabs and anal swabs were basically the same. According to the experimental paired samples to detect EV71, CA16, EV, the result is not consistent with the time of sample collection analysis of enteric viruses in the pathogenesis may be inferred within 2 days of pharyngeal multiply amplification, this time through the possibility of the spread of the respiratory tract.
From July 2010 to December 2011 will be monitoring the clinical diagnosis of HFMD cases of 175 throat swabs and anal swabs, Real-time RT-PCR test results according to the specimen collection time were analyzed for EV71, pharynx swabs in 3 days after the onset, 4-11 days can be detected, and CA16, EV in 3 days after the onset are not positive, EV71 in the respiratory tract with poison duration is longer than CA16 and intestinal virus.
Study on the evolution of enterovirus 71 gene in 3. Jingan District people in Shanghai
In 2010 were randomly selected in different months 19 PCR positive specimens were cultured in Vero cells, including 1 cases of severe culture, 18 non severe cultures of EV71 virus PCR amplified sequences (VPl gene 1200bp) phylogenetic analysis, all 19 strains of EV71 virus in Shandong in 2007 and 2008 Anhui Fuyang EV71 isolates belong to the C genotype in C4a subtype.
The study of the 19 strains of VPl gene of EV71 isolates and representatives of retrieved EV71 genotypes and subtypes from strains of GenBank in the series, including A, B, Cl, C2, C3, C4 and C5 subtype analysis of amino acid and nucleotide sequence, which subtype C4 is the choice of Fuyang in 2008 Anhui city epidemic strains. Analysis showed that 19 strains of VP1 genes and C genes of EV71 isolate type strains nucleotide homology in 84.3%-97.1%, amino acid homology in 88.6%-98.3%; and C4 genotype was most similar to that of the nucleotide homology in amino acid homology in 95.1%-98.3%. 95.0%-97.1%.
Our country since 1998 from Guangdong Province, isolation and identification of C4b subtype EV71 isolates, to 2003 domestic EV71 subtype has been C4b.2003-2004, EV71 C4a and C4b two subtypes in China such as Zhejiang, Guangdong, the coexistence of Taiwan. Since 2005, Shanghai, Henan, Yunnan, Beijing, Shandong. Anhui, Zhejiang and other places of the isolated EV71 were C4a subtype, severe and sporadic cases of the same genotype had no difference. And the United States, Australia, Malaysia, Singapore, Japan and other countries in different period had different EV71 subtypes.
The effect of 4. chlorine containing disinfectant on inactivation of enterovirus type 71
Enterovirus 71 is mainly through the fecal oral route and contact transmission. At present, the Shanghai area preschools are usually used for toys available chlorine concentration 500mg/L of chlorine containing disinfectant on children, towels and desks, disinfection door handles and other surfaces. Therefore, we observed the effect of the concentration of chlorine containing disinfectant in killing in suspension EV71 the virus from the test results. Because of the toxicity of disinfectant on cells, this experiment found the effective chlorine concentration 5min chlorine disinfectant effect of 500mg/L can be reached at least 99.99% of the killing rate of suspension of EV71 virus; and the effective chlorine concentration for the killing rate of 1000mg/L chlorine disinfectant is consistent. Because the test used for virus suspension liquid, disinfectant and intestinal virus can directly contact, but in the daily work of disinfection, chlorine disinfectants to wrapped in a towel, clothes in the killing effect by virus Organic items, effects of electrostatic adsorption and other factors. Therefore, for suspected HFMD contact with contaminated items, by increasing the concentration of effective chlorine (1000mg/L or more) or to extend the time of disinfectant disinfection, ensure the virus inactivation effect, prevent the spread of foot and mouth disease.

【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.1;R181.3

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