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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/72907完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄭謙仁(Chian-Ren Jeng) | |
| dc.contributor.author | Yen-Wen Chen | en |
| dc.contributor.author | 陳彥彣 | zh_TW |
| dc.date.accessioned | 2021-06-17T07:09:57Z | - |
| dc.date.available | 2020-08-18 | |
| dc.date.copyright | 2019-08-18 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-07-22 | |
| dc.identifier.citation | Albina, E., Carrat, C., Charley, B., 1998. Interferon-alpha response to swine arterivirus (PoAV), the porcine reproductive and respiratory syndrome virus. J Interf Cytok Res 18, 485-490.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/72907 | - |
| dc.description.abstract | 豬生殖與呼吸道綜合症（PRRS）對全球養豬產業具有重大經濟影響，主要造成母豬流產、死產、產下木乃伊胎及弱仔等繁殖障礙，以及對生長期豬隻造成呼吸道症狀及生長性能低下。豬生殖與呼吸道綜合症病毒（PRRSV）是具有封套的正鏈單股RNA病毒，分類上屬於動脈病毒科（Arteriviridae）和Porartevirus病毒屬。PRRSV的基因體全長約15kb，並由10個開放閱讀區（open reading frames）所組成，其中開放閱讀區5（ORF5）及其所編碼之封套蛋白GP5是具高度變異性的區域。本研究之實驗目的為深入了解臺灣2014至2019年於田間流行之PRRSV其OFR5序列的基因變異。親緣分析結果顯示，此時期所收集的臺灣PRRSV（n = 73）皆屬於北美型病毒，這些病毒株具有顯著的基因特異性，屬於第三譜系（lineage 3）的北美型PRRSV，並可進一步將所有第三譜系的台灣PRRSV區分為7個不同的亞譜系（sublineage A-G）。台灣PRRSV與其他國家的病毒株和疫苗毒株具有高度基因序列差異，尤其在免疫相關之抗原決定位和N-醣基化位點具有顯著變異。在本研究中所分析的SW2018001-YL病毒株為來自雲林縣的臺灣PRRSV分離株，以次世代定序分析獲得全長基因體，並以RDP重組分析軟體測得在第1-538及14367-末端核苷酸位點發生序列重組，而其親代病毒株可能為HC120904-CHYL病毒株。
臺灣田間豬場自1990年代末期開始使用PRRS商品化活毒減毒疫苗，然而，至今仍不斷有偶發的PRRS疫情傳出。本研究擇一位於新北市之PRRS陽性豬場，進行活毒減毒疫苗之免疫效力評估。該豬場於2017年7月中由A品牌活毒疫苗更換為B品牌活毒疫苗，評估參數包含疫苗毒ORF5序列和田間病毒之相似性、母豬之繁殖性能、血清中之PRRSV含量及ELISA抗體力價等。結果顯示在資料收集期間（2016年9月至2019年2月），同時於分娩後母豬及一週齡仔豬使用B品牌活毒疫苗，能夠有效降低死產率、減少保育豬之病毒血症及增加ELISA血清抗體力價。此外，相較於A品牌活毒疫苗，B品牌活毒疫苗與野外毒株（SW2017001-TP病毒株）在ORF5（GP5）序列及T1抗原決定位具有較高之基因同質性。 | zh_TW |
| dc.description.abstract | Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically significant diseases of swine industry worldwide. It causes reproductive syndrome in sows and respiratory syndrome with poor growth performance in growing pigs. PRRS virus is an enveloped, single-stranded positive-sense RNA virus belonging to the family of Arteriviridae, and the genus of Porartevirus. Within the genome of PRRSV, the ORF5 gene, encoding the major envelope protein, GP5, is the highly variable region. The aim of this study was to gain insight into the genetic variation of the ORF5 gene of PRRSVs circulating in Taiwan from 2014 to 2019. The phylogenetic analysis results showed that all of the field PRRSVs (n = 73) belonged to the North American genotype and have established significant genetic characteristics. These Taiwanese field PRRSVs belong to lineage 3, and they can be further classified into 7 sublineages (sublineage A to G). The Taiwanese PRRSVs show genetically distinct from other global PRRSVs or vaccine viruses, especially in the immunodominant epitopes and the N-glycosylation sites. A Taiwanese PRRSV isolate from Yunlin County, SW2018001-YL strain, was submitted for the next-generation sequencing to obtain the full-length PRRSV genome. Recombinant events were detected by RDP program at the nucleotide positions, 1-538 and 14367-end, and its putative parental strain was HC120904-CHYL.
Although the modified-live virus (MLV) vaccine of PRRSV has been used in Taiwan since the late 1990s, sporadic outbreaks of PRRS still continue occurring. In the present study, a PRRS-endemic pig farm located in New Taipei City was selected for evaluating the efficacy of commercial PRRS MLV vaccines. This pig farm had switched from the brand A PRRS MLV vaccine to the brand B PRRS MLV vaccine since mid-July of 2017. Four parameters, including ORF5 sequence similarity among field and vaccine viruses, reproductive performance, PRRSV viral load in serum, and ELISA anti-PRRSV serum antibody titers, were used to evaluate the efficacy of these two vaccines. The results showed that vaccination of postpartum sows and one-week-old piglets with the brand B PRRS MLV vaccine effectively reduced the stillbirth rate and PRRSV viremia of nursery-growing pigs, and increased the ELISA anti-PRRSV serum antibody titers during the period of September of 2016 to February of 2019. Besides, comparing to the brand A PRRS MLV vaccine strain, the brand B PRRS MLV vaccine strain was more homologous to the field virus (SW2017001-TP strain) in the ORF5 (GP5) and T1 epitope. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T07:09:57Z (GMT). No. of bitstreams: 1 ntu-108-R06644007-1.pdf: 3437527 bytes, checksum: 428716b709f90bd8fb5f59f05a7a10f4 (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | ABSTRACT IN CHINESE............................................................................................i
ABSTRACT IN ENGLISH..........................................................................................iii TABLE OF CONTENTS.............................................................................................. v LIST OF FIGURES…................................................................................................viii LIST OF TABLES…………........................................................................................ix CHAPTER 1. LITERATURE REVIEW 1 1.1 Porcine reproductive and respiratory syndrome (PRRS) 1 1.1.1 Historical background and economic impact on the swine industry 1 1.1.2 Virology of PRRSV 2 1.1.3 Molecular epidemiology of PRRSV 4 1.1.4 Epidemiology, pathogenesis and immunity 5 1.1.5 Clinical signs and pathological changes 10 1.2 Field experiments of PRRS modified-live virus (MLV) vaccines 12 1.2.1 Commercial PRRS MLV vaccines 12 1.2.2 Evaluation the efficacy of PRRS MLV vaccines 15 1.2.2.1 Clinical performance 15 1.2.2.2 PRRSV viremia 16 1.2.2.3 Immune responses to PRRS MLV vaccine 17 CHAPTER 2. MATERIALS AND METHODS 20 2.1 Experimental designs 20 2.1.1 Phylogenetic analysis of the ORF5 gene of PRRSVs 20 2.1.2 Next-generation sequencing of the full PRRSV genome and the analysis of recombination 21 2.1.3 Evaluation of the efficacy of a newly introduced PRRS MLV vaccine in the field 21 2.2 Experimental materials 22 2.2.1 Sample collection 22 2.2.2 Farm history 23 2.3 Experimental methods 23 2.3.1 Phylogenetic analysis of the ORF5 gene of PRRSVs 23 2.3.1.1 Viral RNA extraction 23 2.3.1.2 Real-time multiplex RT-PCR 23 2.3.1.3 RT-PCR 24 2.3.1.4 Cloning and sequencing the full-length ORF5 gene 26 2.3.1.5 Phylogenetic analysis of the ORF5 gene and the encoding GP5 protein 26 2.3.1.6 GenBank accession numbers of ORF5 genes 27 2.3.2 Next-generation sequencing of the full PRRSV genome and the analysis of recombination 27 2.3.2.1 Virus isolation 27 2.3.2.2 Immunocytochemistry staining 28 2.3.2.3 Next-generation sequencing of the full PRRSV genome 29 2.3.2.4 Analysis of recombination 30 2.3.3 Evaluation of the efficacy of a newly introduced PRRS MLV vaccine in the field 30 2.3.3.1 Vaccination program 30 2.3.3.2 Analysis of the ORF5 genes of field virus and PRRS MLV vaccine strains 31 2.3.3.3 Analysis of reproductive performance 31 2.3.3.4 Quantification of PRRSV RNA in serum 31 2.3.3.5 Detection of serum anti-PRRSV antibody titers by ELISA 33 2.3.3.6 Statistical analysis 34 CHAPTER 3. RESULTS 34 3.1 Phylogenetic analysis of the ORF5 gene of PRRSVs 34 3.1.1 RT-PCR survey of clinical samples 34 3.1.2 Phylogenetic analysis of the ORF5 genes 35 3.1.3 Immunodominant epitopes and N-glycosylation sites in GP5 37 3.2 Next-generation sequencing of the full PRRSV genome and analysis of the recombination 40 3.2.1 Virus isolation and immunocytochemistry staining of the SW2018001-YL strain 40 3.2.2 Phylogenetic analysis of the full PRRSV genome of the SW2018001-YL strain………………………………………………….…………………40 3.2.3 Analysis of recombination in the SW2018001-YL strain 41 3.3 Evaluation the efficacy of a newly introduced PRRS MLV vaccine in the field 42 3.3.1 Analysis of the ORF5 gene of the field virus and PRRS MLV vaccines 42 3.3.2 Reproductive performance 43 3.3.3 Quantification of PRRSV RNA in serum 44 3.3.4 ELISA anti-PRRSV serum antibody titer 45 CHAPTER 4. DISCUSSION 45 4.1 Phylogenetic analysis of the ORF5 gene of PRRSVs 45 4.2 Next-generation sequencing of the full PRRSV genome and analysis of recombination 50 4.3 Evaluation the efficacy of a newly introduced PRRS MLV vaccine in the field 53 REFERENCES 89 LIST OF FIGURES Figure 1. Geographical distribution and density of PRRSV ORF5-positive samples 61 2. Phylogenetic tree of the ORF5 gene of type 2 PRRSVs 62 3. Multiple alignments of GP5 amino acid sequences of type 2 PRRSV 63 4. (a) Cytopathic effect of a Taiwanese field PRRSV isolate, SW2018001-YL 64 (b) Immunocytochemistry (ICC) staining result of SW2018001-YL 64 5. Identification of the recombination events in SW2018001-YL strain 65 6. Phylogenetic trees of PRRSVs generated with the nucleotide sequences of full genome using UPGMA parsimony 66 7. Copy numbers of PRRSV RNA in serum 67 8. ELISA anti-PRRSV serum antibody titers 68 LIST OF TABLES Table 1. Lists of primers used in the present study 69 2. Geographical distribution and collecting year of the PRRSV ORF5-positive samples.…………………………………………………………………………70 3. Pairwise inter-sublineage genetic distance comparisons for the ORF5 nucleotide sequence among the lineage 3 PRRSVs detected during 1991-2019 in Taiwan..71 4. Information of the field PRRSV strains circulating in Taiwan during 2014-2019………………………………………………….…………...……………..72 5. The similarity of the ORF5 nucleotide and GP5 amino acid sequences between the Taiwanese field PRRSV strains detected during 2014-2019, MD-001 strain, Ingelvac®, Fostera®, and Amervac® PRRS MLV vaccine strains 77 6. Diverse immunodominant epitopes and putative N-glycosylation sties in GP5 of Taiwanese field PRRSVs detected during 1991-2013 and 2014-2019, VR2332 strain, MD-001 strain, Ingelvac® and Fostera® PRRS MLV vaccine strain…….78 (a) Primary neutralizing epitope (PNE) and decoy epitope (DCE) in GP5 78 (b) T cell epitopes (T1 and T2 epitope) in GP5 80 (c) Putative N-glycosylation sites in GP5 85 7. The phylogenetically closest reference strains of PRRSV and their nucleotide (NT) sequence similarity at each region of the genome to the field strain of SW2018001-YL…..………………………………...…………………………..87 8. Reproductive parameters of sows and newborn piglets in the brand A PRRS MLV vaccinated group (VacA), transition stage group (VacTrans), and brand B PRRS MLV vaccinated group (VacB) in the selected PRRS-endemic farm...………....88 | |
| dc.language.iso | en | |
| dc.title | 臺灣2014-2019年豬生殖與呼吸道綜合症病毒之ORF5序列親緣分析與新商品化活毒疫苗之田間效力評估 | zh_TW |
| dc.title | Phylogenetic analysis of the ORF5 gene of porcine reproductive and respiratory syndrome virus (PRRSV) circulating in Taiwan from 2014 to 2019 and evaluation the efficacy of a newly introduced commercial modified-live PRRSV vaccine in the field | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 張惠雯(Hui-Wen Chang),龐飛(Victor Fei Pang) | |
| dc.contributor.oralexamcommittee | 賈敏原,邱慧英 | |
| dc.subject.keyword | 豬生殖與呼吸道綜合症病毒,開放閱讀區5,親緣分析,次世代定序,重組,活毒減毒疫苗,繁殖性能, | zh_TW |
| dc.subject.keyword | Porcine reproductive and respiratory syndrome virus (PRRSV),Open reading frame 5 (ORF5),Phylogenetic analysis,Next-generation sequencing (NGS),Recombination,Modified-live virus (MLV) vaccine,Reproductive performance, | en |
| dc.relation.page | 103 | |
| dc.identifier.doi | 10.6342/NTU201901660 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2019-07-23 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 分子暨比較病理生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子暨比較病理生物學研究所 | |
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