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研究生:陳雅媚
研究生(外文):Ya-Mei Chen
論文名稱:原位雜合反應顯示豬鐵士古病毒可感染前所未知的豬組織細胞型別
論文名稱(外文):In Situ Hybridization Demonstrates That Porcine Teschovirus Can Infect a Wider Variety of Tissue Cell Types That Has Not Been Revealed
指導教授:王汎熒
口試委員:張伯俊張本恆張家宜李璠
口試日期:2013-07-15
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:獸醫學研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2014
畢業學年度:103
語文別:英文
論文頁數:45
中文關鍵詞:豬鐵士古病毒原位雜合反應
外文關鍵詞:porcine teschovirusISHin situ hybridization
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豬鐵士古病毒(porcine teschoviruses, PTVs)為單股正向RNA病毒,分類為teschovirus屬picornaviridae科。PTV感染會造成多種豬隻疾病;包括非化膿性灰質腦脊髓炎、肺炎、繁殖障礙、腸炎等。PTV在台灣首次於2000年分離出來,目前台灣豬場呈現PTV高度汙染且廣泛分布。本研究目的為探討在地方性感染的豬隻中PTV所可能扮演的致病角色。本實驗室先前收集29頭淘汰豬隻不同臟器並藉巢式聚合酶鏈鎖反應(nested RT-PCR)檢測病毒分布。96.7% 豬隻呈現 PTV陽性,臟器中病毒量最高的為腸道、其次為淋巴器官、腦及內臟。統計顯示,在腦的後段之非化膿性腦炎的出現與 PTV感染具有相關性 (P = 0.054)。本研究則利用原位雜合反應(in situ hybridization, ISH)染色偵測PTV在組織器官中分布的位置。結果顯示PTV廣泛分布於神經系統、淋巴系統、腎臟、空腸、迴腸及結腸內,ISH可偵測到PTV廣泛分佈之前未知的細胞型別,使我們對PTV的致病性有進一步的了解。本研究建立偵測豬鐵士古病毒RNA之原位雜合染色技術並藉以確認病毒的細胞親和性。

Porcine teschoviruses (PTVs) are RNA viruses that are classified within the genus Teschovirus of the family Picornaviridae and cause non-suppurative meningoencephalitis and polioencephalomyelitis. In Taiwan, the PTV infection is the first or secondary swine viral diseases and usually isolated together with other common pathogens. To investigate PTV distribution, tissues were collected from 29 culled postweaning pigs, and the positive rate of PTV detection by nested RT-PCR was 96.7% (by heads). The most positive tissues were the intestines where ileum had the significantly highest detection rate, followed by lymphoid organs, cranial portion of the brain, and visceral organs. Statistics showed that nonsuppurative encephalitis in the caudal brain had an obvious correlation with PTV detection. To recognize the relation between lesions and PTV locations, in situ hybridization (ISH) was performed. PTV positive signals were presented in the nervous system, immune system, kidney, jejunum, ileum and colon. PTVs are able to infect a wider variety of cell types than what has been known. This study furthers our understanding on the pathogenicity and pathogenesis of PTV. The study establishes in situ hybridization technique for PTVs detection and confirms the PTV cell tropism.

中文摘要 I
Abstract II
contents III
List of Tables VI
List of Figures VII
Chapter Ⅰ 1
INTRODUCTION 1
Chapter Ⅱ 3
LITERATURE REVIEW 3
2.1 Porcine teschoviruses (PTVs) 3
2.2 Epidemiology of PTVs 6
2.2.1 Epidemiology in world 6
2.2.2 Epidemiology in Taiwan 6
2.3 Clinical signs of PTV infection 8
2.4 Lesions of PTVs 10
2.4.1 Polioencephalomyelitis 10
2.4.2 Other lesions 12
2.5 Pathogenesis of PTVs 12
2.5.1 Intracellular replication site of PTVs 13
2.6 PTVs detection in tissues morphologically 14
2.6.1 Nervous system 15
2.6.2 Urinary and circulatory systems 15
2.6.3 Respiratory system 16
2.6.4 Immune system 16
2.6.5 Digestive system 16
2.7 In situ hybridization (ISH) 19
2.7.1 Locked nucleic acid (LNA) probe 20
Chapter Ⅲ 20
MATERIALS AND METHODS 20
3.1 Sample preparations 20
3.2 Probe for PTV 21
3.3 In situ hybridization 21
3.3.1 Dewax and hydration 21
3.3.2 Proteinase K digestion and post-fixation 22
3.3.3 Dehydration 22
3.3.4 Hybridization 22
3.3.5 Post hybridization wash 23
3.3.6 Blocking 23
3.3.7 Anti-DIG-AP 23
3.3.8 Detection 23
Chapter Ⅳ 24
RESULTS 24
4.1 Histological examination of HE stained tissue sections 24
4.1.1 Nervous system 24
4.1.2 Urinary and circulatory systems 24
4.1.3 Respiratory system 25
4.1.4 Immune system 25
4.1.5 Digestive system 25
4.2 Examination of in situ hybridization stained tissues sections 26
4.2.1 Distribution of positive cells in each organ 26
Chapter Ⅴ 28
Discussion and Conclusion 28
References 41
Appendix 45


Anonymous. Teschovirus encephalomyelitis (previously enterovirus encephalomyelitis or Teschen/Talfan disease). In: Chapter 2.8.10 Manual of diagnostic tests and vaccines for terrestrial animals, World Organisation for Animal Health, 2008.
Alexander TJL, Richards WPC, Roe CK. An encephalomyelitis of suckling pigs in Ontario. Canadian journal of comparative medicine and veterinary science. 1959;23(10):316-319.
Auerbach J, Prager D, Neuhaus S, Loss U, Witte KH. Grouping of porcine enteroviruses by indirect immunofluorescence and description of two new serotypes. Journal of Veterinary Medicine, Series B. 1994;41(4):277-282.
Boros A, Nemes C, Pankovics P, Kapusinszky B, Delwart E, Reuter G. Porcine teschovirus in wild boars in Hungary. Arch Virol. 2012;157(8):1573-1578.
Chaproniere DM, Done JT, Andrewes CH. Comparative serological studies on Talfan and Teschen diseases and similar conditions. British journal of experimental pathology. 1958;39(1):74-77.
Chiu KC. Detection of porcine teschovirus from culled postweaner pigs. Master degree thesis. National Tainwan University, Taipei, 2010.
Chiu SC, Hua SC, Changc CC, Changa CY, Huanga CC, Pang VF, Wang FI. The role of porcine teschovirus in causing diseases in endemically infected pigs. Veterinary Microbiology. 2012;161(1-2):88-95.
Deng MY, Millien M, Jacques-Simon R, Flanagan J.K., et al. Diagnosis of porcine teschovirus encephalomyelitis in the Republic of Haiti. J Vet Diagn Invest. 2012;24(4):671-678.
Derbyshire, JB, Jessett, DM. Multiplication of some porcine enteroviruses in baby hamster and pig kidney cell lines. Journal of Comparative Pathology. 1967;77(3):237-243.
Dunne HW, Gobble JL, Hokanson JF, Kradel DC, Bubash GR. Porcine reproductive failure associated with a newly identified" SMEDI" group of picorna viruses. American journal of veterinary research. 1965;26(115):1284-1297.
Edington N, Christofinis GJ, Betts AO. Pathogenicity of Talfan and Konratice strains of Teschen virus in gnotobiotic pigs. J Comp Pathol. 1972;82(4):393-399.
Greig AS. Tissue culture studies and electron microscopy of the virus of Teschen disease. Canadian journal of comparative medicine and veterinary science. 1961;25(1):3-7.
Greig AS, Bannister GL, Mitchell D, Corner AH. Studies on pathogenic porcine enteroviruses: II. isolation of virus in tissue culture from brain and feces of clinical cases. Canadian journal of comparative medicine and veterinary science. 1961;25(6):142-148.
Holman JE, Koestner A, Kasza L. Histopathogenesis of porcine polioencephalomyelitis in the germ free pig. Pathologia Veterinaria Online. 1966;3(6):633-651.
Horstmann DM, Manuelidis E, Sprinz H. Neuropathology of teschen disease (virus encephalomyelitis of swine). Society for experimental biology and medicine. 1951;77(1):8-13.
Hu SC. Detection of porcine teschovirus and serotyping of the PTVs in culled post-weaned pigs, and development of qRT-PCR to quantitate PTVs load in tissues. Master degree thesis. National Taiwan University, Taipei, 2011.
Huang J, Gentry RF, Zarkower A. Experimental infection of pregnant sows with porcine enteroviruses. Am J Vet Res. 1980;41(4):469-473.
Huang TS, Wang C, Deng MC, Jeng JJ, Lee SH, Pan CS, Lee MC, Jong MH. The results of virus isolation in swine tissue samples submitted by LDCC and tested by viral isolation and PCR and/or RT-PCR in 2008. Exp. Rep. Animal Health Research Institute. 2009;44: 35-46.
Huck RA, Cartwright SF, Yamanouchi K, Bankowski RA, Howarth JA. Polioencephalomyelitis of pigs – the identification of viruses related to the teschen and T80 groups in the united states. American journal of veterinary research. 1963;24:1207-1211.
Izawa H, Bankowski RA, Howarth JA. Porcine enteroviruses. I. Properties of three isolates from swine with diarrhea and one from apparently normal swine. American journal of veterinary research. 1962;23:1131-1141.
Izawa H, Howarth JA, Bankowski RA. Porcine enteroviruses. II. Pathogenesis of viral agents isolated from the intestinal tract of swine. American journal of veterinary research. 1962;23:1142-1149.
Jiménez-Clavero MA, Fernández C, Ortiz JA, Pro J, Carbonell G, Tarazona JV, Ley V. Teschoviruses as indicators of porcine fecal contamination of surface water. Applied and environmental microbiology. 2003;69(10):6311-6315.
Kasza L. Swine polioencephalomyelitis vieuses isolated from the brains and intestines of pigs. American journal of veterinary research. 1965;26:131-137.
Klobouk A. Encephalomyelitis enzootica suum. Zverolekarsky obzor. 1931;24:436-48.
Koestner A, Kasza L, Holman JE. Electron microscopic evaluation of the pathogenesis of porcine polioencephalomyelitis. Am J Pathol. 1966;49(2):325-337.
Knowles NJ. Porcine enteric picornaviruses, In: Diseases of swine 9th. Blackwell, Iowa. 2006;337-343.
Knowles NJ, Buckley LS, Pereira HG. Classification of porcine enteroviruses by antigenic analysis and cytopathic effects in tissue culture: description of 3 new serotypes. Archives of virology. 1979;62(3):201-208.
L''Ecuyer C, Greig AS. Serological and biological studies on porcine enteroviruses isolated in Canada. The Canadian Veterinary Journal. 1966;7(7):148-154.
Lin YC. Detection of Porcine Teschovirus in Tissues of Experimentally Infected Pigs. Master degree thesis. National Taiwan University, Taipei. 2009.
Long JF, Koestner A, Kasza L. Infectivity of three porcine polioencephalomyelitis viruses for germfree and pathogen-free pigs. American journal of veterinary research. 1966;27(116):274-279.
Long JF, Kasza L, Koestner A. Pericarditis and myocarditis in germfree and colostrum-deprived pigs experimentally infected with a porcine polioencephalomyelitis virus. Journal of Infectious Diseases. 1969;120(2):245-249.
Lou M. In situ hybridization. Acta Histochemica et Cytochemica. 1990;23:1-16.
Manuelidis EE, Sprinz H, Horstmann DM. Pathology of Teschen disease (virus enceph-alomyelitis of swine). Am J Pathol. 1954;30(3):567-597.
Meyer RC, Greider MH, Bohl EH. Electron microscopy of a pathogenic porcine enterovirus. Virology. 1964;22(1):163-165.
Mitchell D, Corner AH, Bannister GL, Greig AS. Studies on Pathogenic Porcine Enteroviruses: 1. Preliminary Investigations. Canadian journal of comparative medicine and veterinary science. 1961;25(4):85-93.
Pogranichniy RM, Janke BH, Gillespie TG, Yoon K-J. A prolonged outbreak of polio-encephalomyelitis due to infection with a group I porcine enterovirus. J Vet Diagn Invest. 2003;15(2):191-194.
Salles MWS, Scholes SFE, Dauber M, Strebelow G, Wojnarowicz C, Hassard L, Acton AC, Bollinger TK. Porcine teschovirus polioencephalomyelitis in western Canada. J Vet Diagn Invest. 2011;23(2):367-373.
Singh SK, Koshkin AA, Wengel J, Nielsen, P. LNA (locked nucleic acids): synthesis and high-affinity nucleic acid recognition. Chemical Communications. 1998;4:455-456.
Smith TJ, Baker T. Picornaviruses: epitopes, canyons, and pockets. Adv Virus Res. 1999;52:1-23.
Takahashi M, Seimiya YM, Seki Y, Yamada M. A piglet with concurrent polioencephalomyelitis due to porcine teschovirus and postweaning multisystemic wasting syndrome. J Vet Med Sci. 2008;70(5):497-500.
Watanabe H, Pospisil Z, Mensik J. Study on the pigs infected with virulent teschen disease virus (KNM strain) with special redernce to immunofluorescence. Japanese Journal of Veterinary Research. 1971;19(4):87-102.
Yamada M, Kaku Y, Nakamura K, Yoshii M, Yamamoto Y, Miyazaki A, Tsunemitsu H, Narita M. Immunohistochemical detection of porcine teschovirus antigen in the formalin-fixed paraffin-embedded specimens from pigs experimentally infected with porcine teschovirus. J Vet Med A Physiol Pathol Clin Med. 2007;54(10):571-574.
Yamada M, Kozakura R, Ikegami R, Nakamura K, Kaku Y, Yoshii M, Haritani M. En-terovirus encephalomyelitis in pigs in Japan caused by porcine teschovirus. Vet Rec. 2004;155(10):304-306.
Yamada M, Kozakura R, Kaku Y, Nakamura K, Yamamoto Y, Yoshii M, Miyazaki A, Tsunemitsu H, Narita M. Immunohistochemical distribution of viral antigens in pigs naturally infected with porcine teschovirus. J Vet Med Sci. 2008;70(3):305-308.
Yamada M, Kozakura R, Nakamura K, Yamamoto Y, Yoshii M, Kaku Y, Miyazaki A, Tsunemitsu H, Narita M. Pathological Changes in Pigs Experimentally Infected with Porcine Teschovirus. J Comp Path. 2009;141(4):223-228.
Zell R, Krumbholz A, Henke A, Birch-Hirschfeld E, Stelzner A, Doherty M, Wurm R. Detection of porcine enteroviruses by nRT–PCR: differentiation of CPE groups I–III with specific primer sets. Journal of virological methods. 2000;88(2):205-218.


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