福州市手足口病病原調(diào)查及基因進化研究
發(fā)布時間：2018-07-27 14:46
【摘要】：目的 1、對2012年福州市手足口病進行病原學調(diào)查，旨在全面了解福州地區(qū)手足口病的腸道病毒譜，分析主要病毒型別的遺傳進化特征，為手足口病的預防控制提供科學依據(jù)。 2、鑒定2012年福州市CVA6陽性病例的比例，分析CVA6陽性病例的流行病學特征。 方法 1、收集2012年福州市區(qū)臨床表現(xiàn)為手足口病的樣本1031份，通過熒光定量RT-PCR、巢式RT-PCR以及NCBI提供的BLAST序列比對確定腸道病毒型別。 2、利用CVA6特異性引物鑒定2012年福州市其他腸道病毒，分析CVA6陽性病例的三間分布。 3、選擇2012年的EV71、CVA16、CVA6陽性樣本進行病毒分離和核酸提取，通過RT-PCR對其進行VP1和5'-UTR序列擴增，序列測定后使用mega5.02進行基因片段的拼接、比對和分析，并將福州株與GenBank上相關的代表株進行同源性和親緣進化分析。 結果 1、1031份標本中878份EV陽性，檢出率為85.16%（878/1031），其中EV71426例，，陽性率為45.52%(426/878)；CVA1693例，陽性率為10.60%(93/878)；其他腸道病毒359例，陽性率為40.88%(359/878)，其中CVA6為218例，陽性率為24.83%（218/878）。此外，還發(fā)現(xiàn)CVA10和Echo3腸道病毒。 2、流行病學特征。時間上：CVA6構成比情況在各月有較大差異，總體起伏明顯。2012年9月的病例多數(shù)由其他腸道病毒引起，而且以CVA6為主。該月病例中CVA6型占62.50%。地區(qū)上：A6分布存在地區(qū)差異。永泰縣明顯高于全市其它縣（市）區(qū)，A6陽性率為54.55%（95%CI:25.12%-83.97%）。人群間：0-5歲年齡組兒童為主要發(fā)病人群（構成比為94.04%）。男女性別比為1.66:1。 3、福州EV71株與C4a亞型毒株具有最高的VP1同源性，為92.9%-97.3%，與2008年安徽阜陽毒株同源性為96.7%。VP1進化樹顯示：EV71可以劃分為A、B、C三個基因型，福州株屬于C4a亞型。在5'-UTR核苷酸序列方面，福州EV71分離株和C4a亞型代表株具有最高的核苷酸序列同源性，為96.7%-98.8%。5'-UTR進化樹顯示，福州地方株與C4a亞型的眾多毒株聚集成簇。 4、福州CVA16地方株和B1b亞型代表株具有最高的VP1核苷酸序列同源性，為90.8%-93.2%。VP1進化樹顯示，CVA16可以劃分為A、B兩個基因型，福州地方株屬于B1b亞型。在5'-UTR核苷酸序列方面，福州CVA16分離株和B1b亞型代表株具有最高的核苷酸序列同源性，為94.8%-97%。5'-UTR進化樹顯示：福州地方株與B1b亞型的眾多毒株聚集成簇。 5、CVA6福州地方株與其原型株(Gdula株)的VP1核苷酸序列同源性為77.4%-79.8%。VP1進化樹顯示，CVA6可以劃分為A、B、C三個簇（cluster），CVA6福州地方株屬于Cluster C，福州CVA16分離株和C簇代表株具有最高的核苷酸序列同源性，為86.1%-98%。在5'-UTR核苷酸序列方面，CVA6福州地方株與A、C各簇分離株的核苷酸序列的同源性依次為89.1%-91.3%和91.7%-99.7%。福州CVA16分離株和C簇代表株具有最高的核苷酸序列同源性。5'-UTR進化樹顯示，基于5'-UTR和VP1基因的樹形在形態(tài)上接近。 結論 1、EV71、CVA16和CVA6是福州市HFMD的主要腸道病原體。CVA6病毒已經(jīng)超過CVA16成為第二大病原體。 2、2012年福州EV71型屬C4a亞型，CVA16為B1b亞型，CVA6屬于C Cluster，與近幾年中國大陸各基因型優(yōu)勢株流行趨勢基本一致。 3、福州地區(qū)手足口病腸道病毒的三個型別（EV71、CVA16、CVA6）在基于VP1和5'UTR區(qū)的進化樹的形態(tài)接近。綜合運用這兩種進化樹可能有利于臨床診斷和疾病防控。
[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.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R512.5
本文編號：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.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R512.5
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