福州市手足口病病原调查及基因进化研究
发布时间：2018-07-27 14:46
【摘要】：目的 1、对2012年福州市手足口病进行病原学调查，旨在全面了解福州地区手足口病的肠道病毒谱，分析主要病毒型别的遗传进化特征，为手足口病的预防控制提供科学依据。 2、鉴定2012年福州市CVA6阳性病例的比例，分析CVA6阳性病例的流行病学特征。 方法 1、收集2012年福州市区临床表现为手足口病的样本1031份，通过荧光定量RT-PCR、巢式RT-PCR以及NCBI提供的BLAST序列比对确定肠道病毒型别。 2、利用CVA6特异性引物鉴定2012年福州市其他肠道病毒，分析CVA6阳性病例的三间分布。 3、选择2012年的EV71、CVA16、CVA6阳性样本进行病毒分离和核酸提取，通过RT-PCR对其进行VP1和5'-UTR序列扩增，序列测定后使用mega5.02进行基因片段的拼接、比对和分析，并将福州株与GenBank上相关的代表株进行同源性和亲缘进化分析。 结果 1、1031份标本中878份EV阳性，检出率为85.16%（878/1031），其中EV71426例，，阳性率为45.52%(426/878)；CVA1693例，阳性率为10.60%(93/878)；其他肠道病毒359例，阳性率为40.88%(359/878)，其中CVA6为218例，阳性率为24.83%（218/878）。此外，还发现CVA10和Echo3肠道病毒。 2、流行病学特征。时间上：CVA6构成比情况在各月有较大差异，总体起伏明显。2012年9月的病例多数由其他肠道病毒引起，而且以CVA6为主。该月病例中CVA6型占62.50%。地区上：A6分布存在地区差异。永泰县明显高于全市其它县（市）区，A6阳性率为54.55%（95%CI:25.12%-83.97%）。人群间：0-5岁年龄组儿童为主要发病人群（构成比为94.04%）。男女性别比为1.66:1。 3、福州EV71株与C4a亚型毒株具有最高的VP1同源性，为92.9%-97.3%，与2008年安徽阜阳毒株同源性为96.7%。VP1进化树显示：EV71可以划分为A、B、C三个基因型，福州株属于C4a亚型。在5'-UTR核苷酸序列方面，福州EV71分离株和C4a亚型代表株具有最高的核苷酸序列同源性，为96.7%-98.8%。5'-UTR进化树显示，福州地方株与C4a亚型的众多毒株聚集成簇。 4、福州CVA16地方株和B1b亚型代表株具有最高的VP1核苷酸序列同源性，为90.8%-93.2%。VP1进化树显示，CVA16可以划分为A、B两个基因型，福州地方株属于B1b亚型。在5'-UTR核苷酸序列方面，福州CVA16分离株和B1b亚型代表株具有最高的核苷酸序列同源性，为94.8%-97%。5'-UTR进化树显示：福州地方株与B1b亚型的众多毒株聚集成簇。 5、CVA6福州地方株与其原型株(Gdula株)的VP1核苷酸序列同源性为77.4%-79.8%。VP1进化树显示，CVA6可以划分为A、B、C三个簇（cluster），CVA6福州地方株属于Cluster C，福州CVA16分离株和C簇代表株具有最高的核苷酸序列同源性，为86.1%-98%。在5'-UTR核苷酸序列方面，CVA6福州地方株与A、C各簇分离株的核苷酸序列的同源性依次为89.1%-91.3%和91.7%-99.7%。福州CVA16分离株和C簇代表株具有最高的核苷酸序列同源性。5'-UTR进化树显示，基于5'-UTR和VP1基因的树形在形态上接近。 结论 1、EV71、CVA16和CVA6是福州市HFMD的主要肠道病原体。CVA6病毒已经超过CVA16成为第二大病原体。 2、2012年福州EV71型属C4a亚型，CVA16为B1b亚型，CVA6属于C Cluster，与近几年中国大陆各基因型优势株流行趋势基本一致。 3、福州地区手足口病肠道病毒的三个型别（EV71、CVA16、CVA6）在基于VP1和5'UTR区的进化树的形态接近。综合运用这两种进化树可能有利于临床诊断和疾病防控。
[Abstract]:objective
1, in 2012, the etiology of hand foot and mouth disease in Fuzhou was investigated. The purpose of this study was to understand the enterovirus spectrum of hand foot and mouth disease in Fuzhou and analyze the characteristics of the genetic evolution of the main virus types, so as to provide scientific basis for the prevention and control of hand foot and mouth disease.
2, we identified the proportion of CVA6 positive cases in Fuzhou in 2012 and analyzed the epidemiological characteristics of CVA6 positive cases.
Method
1, 1031 samples of hand foot and mouth disease (HFMD) in Fuzhou city in 2012 were collected, and the comparison of BLAST sequences provided by fluorescent quantitative RT-PCR, nested RT-PCR and NCBI was used to determine the type of enterovirus.
2, the CVA6 specific primers were used to identify other enterovirus in Fuzhou in 2012, and the distribution of three positive CVA6 cases was analyzed.
3, the EV71, CVA16 and CVA6 positive samples in 2012 were selected for virus isolation and nucleic acid extraction, and VP1 and 5'-UTR sequences were amplified by RT-PCR. After sequencing, the sequences were spliced, compared and analyzed using mega5.02, and the homology and phylogenetic evolution of the Fuzhou strain related to GenBank were analyzed.
Result
Of the 11031 specimens, 878 EV were positive, the detection rate was 85.16% (878/1031), and the positive rate was 45.52% (426/878) in EV71426 cases; the positive rate was 10.60% (93/878) in CVA1693 cases and 359 of the other enterovirus cases, the positive rate was 40.88% (359/878), of which CVA6 was 218 and the positive rate was 24.83% (218/878). Furthermore, CVA10 and Echo3 enterovirus was found.
2, epidemiological characteristics. Time: CVA6 composition ratio has great difference in each month. The overall fluctuation is obvious in September of.2012 years, most of the cases are caused by other enteroviruses and are mainly CVA6. In this month, the CVA6 type in 62.50%. area: A6 distribution exists regional difference. Yongtai county is obviously higher than the other county (city) area of the city, A6 positive rate. It was 54.55% (95%CI:25.12%-83.97%). Among the children: the 0-5 year old group was the main group (the constituent ratio was 94.04%). The ratio of male to female was 1.66:1.
3, the Fuzhou EV71 strain and the C4a subtype have the highest VP1 homology, which is 92.9%-97.3%. The homology of the Fuyang strain in Anhui in 2008 shows that the EV71 can be divided into A, B, C, and the Fuzhou strain belongs to the C4a subtype. In the 5'-UTR nucleotide sequence, the Fuzhou EV71 isolates and the representative strains of the subtype are the highest nuclei. The nucleotide sequence homology of the 96.7%-98.8%.5'-UTR phylogenetic tree showed that many strains of Fuzhou local strains and C4a subtypes were clustered together.
4, Fuzhou CVA16 local strain and B1b subtype represent the highest VP1 nucleotide sequence homology. For 90.8%-93.2%.VP1 evolution tree, CVA16 can be divided into A, B two genotypes, and Fuzhou local strains belong to B1b subtype. In 5'-UTR nucleotide sequence, the Fuzhou CVA16 isolates and B1b subtypes have the highest nucleotide sequence homology The 94.8%-97%.5'-UTR evolution tree shows that many strains of Fuzhou local strains and B1b subtypes are clustered together.
5, the homology of the VP1 nucleotide sequence of CVA6 Fuzhou local strain and its prototype strain (Gdula strain) shows that the CVA6 can be divided into A, B, C three clusters (cluster), CVA6 Fuzhou local strains belong to Cluster C, and the Fuzhou isolated strain and the representative plant have the highest nucleotide sequence homology. On the other hand, the nucleotide sequences of CVA6 Fuzhou local strains and A, C cluster isolates were homologous to 89.1%-91.3% and 91.7%-99.7%. Fuzhou CVA16 isolates and C cluster strains, which showed the highest nucleotide sequence homologous.5'-UTR evolution tree, and the tree shape based on 5'-UTR and VP1 genes was similar in morphology.
conclusion
1, EV71, CVA16 and CVA6 are the main enteric pathogens of HFMD in Fuzhou. The.CVA6 virus has exceeded CVA16 and has become the second largest pathogen.
In the 22012 year, Fuzhou EV71 belongs to C4a subtype, CVA16 is B1b subtype and CVA6 belongs to C Cluster, which is basically consistent with the prevalent trend of dominant genotype in mainland China in recent years.
3, the three types (EV71, CVA16, CVA6) of the hand foot and mouth disease enterovirus (EV71, CVA16, CVA6) are close to the evolution tree based on the VP1 and 5'UTR regions. The comprehensive use of these two evolutionary trees may be beneficial to clinical diagnosis and disease prevention and control.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.5
本文编号：2148157
[Abstract]:objective
1, in 2012, the etiology of hand foot and mouth disease in Fuzhou was investigated. The purpose of this study was to understand the enterovirus spectrum of hand foot and mouth disease in Fuzhou and analyze the characteristics of the genetic evolution of the main virus types, so as to provide scientific basis for the prevention and control of hand foot and mouth disease.
2, we identified the proportion of CVA6 positive cases in Fuzhou in 2012 and analyzed the epidemiological characteristics of CVA6 positive cases.
Method
1, 1031 samples of hand foot and mouth disease (HFMD) in Fuzhou city in 2012 were collected, and the comparison of BLAST sequences provided by fluorescent quantitative RT-PCR, nested RT-PCR and NCBI was used to determine the type of enterovirus.
2, the CVA6 specific primers were used to identify other enterovirus in Fuzhou in 2012, and the distribution of three positive CVA6 cases was analyzed.
3, the EV71, CVA16 and CVA6 positive samples in 2012 were selected for virus isolation and nucleic acid extraction, and VP1 and 5'-UTR sequences were amplified by RT-PCR. After sequencing, the sequences were spliced, compared and analyzed using mega5.02, and the homology and phylogenetic evolution of the Fuzhou strain related to GenBank were analyzed.
Result
Of the 11031 specimens, 878 EV were positive, the detection rate was 85.16% (878/1031), and the positive rate was 45.52% (426/878) in EV71426 cases; the positive rate was 10.60% (93/878) in CVA1693 cases and 359 of the other enterovirus cases, the positive rate was 40.88% (359/878), of which CVA6 was 218 and the positive rate was 24.83% (218/878). Furthermore, CVA10 and Echo3 enterovirus was found.
2, epidemiological characteristics. Time: CVA6 composition ratio has great difference in each month. The overall fluctuation is obvious in September of.2012 years, most of the cases are caused by other enteroviruses and are mainly CVA6. In this month, the CVA6 type in 62.50%. area: A6 distribution exists regional difference. Yongtai county is obviously higher than the other county (city) area of the city, A6 positive rate. It was 54.55% (95%CI:25.12%-83.97%). Among the children: the 0-5 year old group was the main group (the constituent ratio was 94.04%). The ratio of male to female was 1.66:1.
3, the Fuzhou EV71 strain and the C4a subtype have the highest VP1 homology, which is 92.9%-97.3%. The homology of the Fuyang strain in Anhui in 2008 shows that the EV71 can be divided into A, B, C, and the Fuzhou strain belongs to the C4a subtype. In the 5'-UTR nucleotide sequence, the Fuzhou EV71 isolates and the representative strains of the subtype are the highest nuclei. The nucleotide sequence homology of the 96.7%-98.8%.5'-UTR phylogenetic tree showed that many strains of Fuzhou local strains and C4a subtypes were clustered together.
4, Fuzhou CVA16 local strain and B1b subtype represent the highest VP1 nucleotide sequence homology. For 90.8%-93.2%.VP1 evolution tree, CVA16 can be divided into A, B two genotypes, and Fuzhou local strains belong to B1b subtype. In 5'-UTR nucleotide sequence, the Fuzhou CVA16 isolates and B1b subtypes have the highest nucleotide sequence homology The 94.8%-97%.5'-UTR evolution tree shows that many strains of Fuzhou local strains and B1b subtypes are clustered together.
5, the homology of the VP1 nucleotide sequence of CVA6 Fuzhou local strain and its prototype strain (Gdula strain) shows that the CVA6 can be divided into A, B, C three clusters (cluster), CVA6 Fuzhou local strains belong to Cluster C, and the Fuzhou isolated strain and the representative plant have the highest nucleotide sequence homology. On the other hand, the nucleotide sequences of CVA6 Fuzhou local strains and A, C cluster isolates were homologous to 89.1%-91.3% and 91.7%-99.7%. Fuzhou CVA16 isolates and C cluster strains, which showed the highest nucleotide sequence homologous.5'-UTR evolution tree, and the tree shape based on 5'-UTR and VP1 genes was similar in morphology.
conclusion
1, EV71, CVA16 and CVA6 are the main enteric pathogens of HFMD in Fuzhou. The.CVA6 virus has exceeded CVA16 and has become the second largest pathogen.
In the 22012 year, Fuzhou EV71 belongs to C4a subtype, CVA16 is B1b subtype and CVA6 belongs to C Cluster, which is basically consistent with the prevalent trend of dominant genotype in mainland China in recent years.
3, the three types (EV71, CVA16, CVA6) of the hand foot and mouth disease enterovirus (EV71, CVA16, CVA6) are close to the evolution tree based on the VP1 and 5'UTR regions. The comprehensive use of these two evolutionary trees may be beneficial to clinical diagnosis and disease prevention and control.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.5
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