云南省边境地区麻疹流行特征和基因特征分析研究

发布时间：2018-05-31 23:37

本文选题：边境地区 + 麻疹　；参考：《昆明医科大学》2014年硕士论文

【摘要】：目的通过对云南省边境地区2013年麻疹流行病学特征和人群免疫情况的描述和分析,了解边境地区麻疹在不同人群、地区、时间的分布情况和人群免疫状况。通过对边境地区麻疹的分子生物学监测,掌握云南省边境地区麻疹病毒株基因的型别及与代表毒株相关性,为云南省边境地区及全省麻疹防控策略和措施的制定提供科学依据,对推进全省麻疹消除进程具有重要的现实意义。研究方法采用描述性流行病学方法,描述云南省边境地区2013年麻疹流行病学特征和人群免疫情况。采用实验流行病学方法,使用Vero/SLAM细胞(淋巴信号激活因子转染的非洲绿猴肾细胞)进行麻疹病毒的分离培养,用逆转录一聚合酶链反应(RT-PCR)检测方法对病例进行确诊和分离病毒的鉴定,对未分离到病毒的标本直接提取病毒核酸并进行RT-PCR和基因测序。描述性流行病学研究的数据录入使用Epidata3.0软件,录入后导出Excel后进行整理。数据分析利用SPSS17.0软件。实验流行病学研究的数据与WHO代表株N基因羧基末端450个核苷酸序列做比对,用MEGA5.1软件进行序列基因亲缘关系分析,做基因亲缘性进化树。结果云南省边境地区2013年麻疹流行病学特征：(1)本研究共报告麻疹确诊病例141例(含实验室确诊和临床诊断)。(2)年龄分布：发病比例最高的年龄组为20-49岁成人组,占58.86%(83/141),其次是0-1.5岁儿童,占27.66%(39/141)；从年龄别发病率来看,0-1岁组的婴儿年龄别发病率最高68.56/10万(34/49592)。(3)男女性别比例为0.94,但1岁年龄组男女性别比为3.0。(4)各职业人群中,农民发病52例(36.8%),散居儿童发病48例(34.04%),其他职业的发病较少。(5)发病特征：发病有明显的季节性,春夏季为边境地区麻疹的高发季节,发病115例,占全年发病的81.56%,暴发和散发麻疹疫情均有相同的季节性规律。除瑞丽市、陇川县和孟连县外均有麻疹病例发生,高发区主要集中在芒市15.66/.10万(62/395913)、临翔区7.67/10万(25/325945)、思茅区6.07/10万(18/296540)、勐海县4.84/10万(13/268595),均高于全省发病率3.04/10万。(6)免疫史：麻疹病例中仅有23例(16.31%)病例有确切免疫史,“0”剂次麻疹病例有70例(49.65%),免疫剂次不详的麻疹病例有48例(34.04%)。按年龄分布情况为：8月龄18例、≥18岁88例,占75.18%(106/141),为现行免疫策略覆盖外人群；8月龄-6岁共31例,为适龄接种人群,病例中有确切接种史的病例仅有13例(41.94%),“0”剂次麻疹病例有16例(51.61%),免疫史不详2例(6.45%)。(7)麻疹疫苗接种情况：云南省边境地区各县(区)2013年含麻疹组份疫苗的报告接种率均大于98%。云南省边境地区2013年麻疹病毒监测及基因特征：(1)麻疹病毒分离：共收集到393份标本,有19份标本发生CPE,其中尿液标本13份,分离率6.84%,咽拭子标本6份,分离率2.96%。对未分离到病毒株麻疹病例标本提取核酸,共获取30株病毒核酸。(2)RT-PCR结果：PCR产物纯化后,经2%琼脂糖凝胶电泳鉴定,均在700bp处有一明显阳性带。(3)麻疹病毒基因型分布：19株分离到的病毒及30株病毒核酸经基因序列测定和分析,共存在3种基因型,其中HI基因型分布于5个边境县(区),老挝勐赛省南磨县及缅甸贵概；D9基因型分布于德宏州与缅甸接壤的3个县及保山市龙陵县和缅甸勐惹、木姐、贵概；A基因型来源于德宏州芒市。(4)基因亲缘性关系分析：用MEGA5.1软件对病毒分离物和病毒核酸做基因亲缘性关系分析。结果提示,12株为H1基因型的H1a基因亚型(24.49%),36株为D9基因型(73.47%),1株为A基因型(中国沪191疫苗株、2.04%)。(5)36株D9基因型与世界卫生组织D9基因型代表株比较：分为5个组(代表株单独为一组)。各组间核苷酸同源性为96.0%-99.5%,氨基酸同源性为95.2%-99.3%。Custer1和Cluster2仅包含缅籍毒株,Cluster3、Cluster4均由云南省本土分离株和缅籍毒株组成(6)12株H1a亚型和世界卫生组织H1a基因亚型和H1b基因亚型代表株比较：分为4个组(代表株各为一组)。各组间核苷酸同源性为97.1%-99.1%,氨基酸同源性为95.8%～99.3%。Cluster1仅包含1株缅籍毒株,Cluster2由云南省本土分离株和老挝籍毒株组成。(7)D9基因型麻疹疫情暴发的监测：2013年5～8月,德宏州芒市风平镇共发生麻疹暴发疫情3起,涉及病例22例,病例数在2～13例之间；3起暴发疫情共涉及13个自然村(社),其中3个村发生点状暴发(相章村、老光村、芒赛村),采集病原学标本,分离到活病毒3株和检测到5株病毒核酸,合计8株,均为同一起暴发疫情中分离和检测到的D9基因型病毒株。 (8)1株A基因型和世界卫生组织A基因型代表株比较,核苷酸同源性为99.1%,氨基酸同源性为99.3%。结论云南省边境地区2013年麻疹发病比例最高的年龄组为20～49岁成人组(58.86%),其次是0～1.5岁儿童(27.66%)；其中0～1岁的婴儿年龄别发病率最高(68.56/10万)；发病以农民(36.88%)和散居儿童(34.04%)为主,共100例(70.92%)；冬春季为麻疹高发季节；高发区主要集中在德宏州(芒市)、临沧市(临翔区)、普洱市(思茅区)、西双版纳州(勐海县)4个边境州(市)所辖的4个县(区),4个县的发病率均高于全省发病率(3.04/10万)；病例中仅有23例(16.31%)病例有确切免疫史,适龄接种人群病例中有确切免疫史者仅占41.94%(15例),无免疫史者(“0”剂次)占51.61%(16例)。在云南省边境地区,存在H1和D9两种基因型麻疹病毒的循环,D9基因型是由缅甸输入的,现为德宏州的优势基因型,且该基因型在德宏州与缅甸接壤的边境县和缅甸相互传播；H1基因型在老挝和云南省与老挝接壤的边境地区相互传播。A基因型为中国沪191疫苗株相似株。
[Abstract]:objective
Through the description and analysis of the epidemiological characteristics of measles and population immunity in the border area of Yunnan Province in 2013, the distribution of measles in the border areas, the distribution of time and the immune status of the population are understood. The molecular biological monitoring of measles in the border area and the type of the gene of measles virus in the border area of Yunnan province are grasped. It provides scientific basis for the formulation of measles prevention and control strategies and measures in Yunnan border area and province, and has important practical significance for promoting the process of measles elimination in the province.
A descriptive epidemiological method was used to describe the epidemiological characteristics of measles and the population immunity in the border area of Yunnan province in 2013. By using experimental epidemiological methods, the measles virus was isolated and cultured with Vero/SLAM cells (transfected by lymphatic signal activating factor), and a reverse transcription polymerase chain reaction (RT-PCR) was used. The detection method was used to identify and isolate the virus, and directly extract virus nucleic acid from the specimens without virus isolation, and carry out RT-PCR and gene sequencing.
Epidata3.0 software was used for data entry of descriptive epidemiological study. After the input of Excel, the data was analyzed using SPSS17.0 software. The data of the data were compared with the 450 nucleotide sequences of the carboxyl terminal of the WHO representative strain of the N gene, and the genetic relationship of the sequence genes was analyzed by MEGA5.1 software, and the gene parent was used as a gene parent. Phylogenetic tree.
Result
The epidemiological characteristics of measles in Yunnan border area in 2013: (1) 141 cases of measles confirmed cases (including laboratory diagnosis and clinical diagnosis) were reported. (2) age distribution: the age group with the highest incidence was 20-49 year old adult group, 58.86% (83/141), followed by 0-1.5 years, accounting for 27.66% (39/141); from the age incidence rate, 0-1 The age incidence rate of the age group was the highest 68.56/10 million (34/49592). (3) the sex ratio of men and women was 0.94, but among the 1 years old age group, the sex ratio of men and women was 3.0. (4), 52 cases (36.8%), 48 cases (34.04%) of scattered children, and less diseases in other occupations. (5) the onset of the disease was seasonal, and spring and summer was the border. In the area of high incidence of measles in the region, 115 cases were found, accounting for 81.56% of the annual incidence of measles. The outbreak and epidemic of measles had the same seasonal regularity. In addition to Ruili, there were measles cases in Longchuan County and Meng Lian county. The high incidence areas mainly concentrated in Manshi 15.66/.10 million (62/395913), 7.67/10 million (25/325945) in Xiang Xiang District, and 6.07/10 million (18/29654) in Simao district (18/29654) 0), 4.84/10 million (13/268595) in Menghai County, higher than the province's incidence of 3.04/10 million. (6) immunization history: only 23 cases (16.31%) of measles cases have definite immunization history, 70 cases (49.65%) with "0" dose of measles cases, 48 cases of measles (34.04%). According to age distribution, 8 month old 18 cases, 18 years 88 cases, occupy 75.18% (23). 106/141), for the current immunization strategy, 31 cases of 8 month old -6 years old, only 13 cases (41.94%) had a history of inoculation, 16 cases of measles (51.61%), 2 cases of immunization (6.45%). (7) measles vaccination: the measles group in the counties (districts) of Yunnan border area (District) in 2013 The inoculation rate of the vaccine was more than 98%.
Yunnan border area measles virus monitoring and gene characteristics in 2013: (1) measles virus isolation: a total of 393 specimens were collected, 19 specimens were CPE, including 13 urine specimens, 6.84% separation rate, 6 pharynx swabs, and isolation rate 2.96%. to extract nucleic acid from the unisolated measles case specimens. (2) RT- PCR results: after the purification of PCR products, a positive positive band was found at 700bp by 2% agarose gel electrophoresis. (3) the distribution of measles virus genotypes: 19 strains of isolated viruses and 30 strains of virus nucleic acid by gene sequencing and analysis, there were 3 genotypes, of which the HI based type was distributed in 5 border counties (regions), and South Mill County, Meng SiO Province, Laos And Burma Guizhou Province, D9 genotypes were distributed in 3 counties, Dehong and Burma, and Baoshan City, dragon Ling county and Burma Meng Provo, Mu Jie and Guizhou Province. A genotypes were derived from Manshi in Dehong. (4) genetic relationship analysis of the gene: MEGA5.1 software was used to analyze the genetic relationship between virus isolates and virus nucleic acids. The results suggested that 12 strains were H1 based The H1a gene subtype (24.49%), 36 strains were D9 genotypes (73.47%), 1 were A genotype (China Shanghai 191 vaccine strain, 2.04%). (5) 36 D9 genotypes were compared with the WHO D9 genotypes, which were divided into 5 groups (representative strains alone). The homology of nucleoside was 96.0%-99.5%, and the amino acid homology was 95.2%-99.3%.Custer1 And Cluster2 only contained Myanmar strains, Cluster3 and Cluster4 were composed of local isolates and Burmese strains in Yunnan province (6) 12 strains of H1a subtype, WHO H1a gene subtype and H1b gene subtype, which were divided into 4 groups (each group was one group). The homology of nucleoside acid was 97.1%-99.1%, and the amino acid homology was 95.8% ~ 9. 9.3%.Cluster1 contains only 1 strains of Myanmar, Cluster2 is composed of native isolates and Laotian strains in Yunnan. (7) monitoring of the outbreak of D9 genotype measles outbreak: 5~8 months of 2013, 3 cases of measles outbreak in Manshi Ping Town, Dehong, involving 22 cases, 2~13 cases, and 3 outbreaks involving 13 natural cases. The village (Society), of which 3 villages have occurred punctuate outbreaks (phase Zhang village, Lao Guang village, Muang Xay Village), collect pathogenic specimens, separate the live virus 3 strains and detect 5 strains of virus nucleic acid and total 8 strains, all of the D9 genotypic virus strains isolated and detected in the same outbreak.
(8) compared with 1 A genotype and WHO A genotype, nucleotide homology was 99.1%, amino acid homology was 99.3%.
Conclusion the highest incidence rate of measles in Yunnan border area in 2013 is 20~49 year old adult group (58.86%), followed by 0 to 1.5 year old children (27.66%); 0~1 year old infants have the highest incidence of age (68.56/10 million); the incidence of the disease is farmers (36.88%) and scattered children (34.04%), 100 cases (70.92%), and high incidence of measles in winter and spring. Season; high incidence area mainly concentrated in Dehong (Mango), Lincang City (Linxiang District), Pu'er City (Simao District), Xishuangbanna state (Menghai county) 4 border states (districts), the incidence of 4 counties is higher than the incidence of the province (3.04/10 million); only 23 cases (16.31%) cases have a definite immunization history, a case of age inoculation crowd. Only 41.94% (15 cases) with definite immunization history and 51.61% (16 cases) without immunization history ("0"). In Yunnan border area, there are two types of H1 and D9 genotypes of measles virus, the D9 genotype is imported from Burma and now the dominant genotype in Dehong, and the genotypes are in Dehong border Burma border county and Burma. Transmission; H1 genotype was transmitted in Laos and Yunnan borders with Laos;.A genotype was similar to Chinese vaccine strain 191 in Shanghai.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R511.1

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