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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 鄭謙仁 | |
dc.contributor.author | Chun Wang | en |
dc.contributor.author | 王羣 | zh_TW |
dc.date.accessioned | 2021-06-07T23:51:41Z | - |
dc.date.copyright | 2014-01-27 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-01-13 | |
dc.identifier.citation | Alhassan, A., Thekisoe O.M., Yokoyama, N., Inoue, N., Motloang, M.Y., Mbati, P.A., Yin, H., Katayama, Y., Anzai, T., Sugimoto, C., Igarashi, I., 2007. Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis. Veterinary Parasitology 143, 155–160.
Allan, G.M., McNeilly, F., Kennedy, S., Daft, B., Clarke, E.G., Ellis, J.A., Haines, D.M., Meehan, B.M., Adair, B.M., 1998. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. Journal of Veterinary Diagnostic Investigation 10, 3–10. Allan, G.M., McNeilly, F., Cassidy, J.P., Reilly, G.A., Adair, B., Ellis, W,A., McNulty, M.S., 1995. Pathogenesis of porcine circovirus; experimental infections of colostrum deprived piglets and examination of pig foetal material. Veterinary Microbiology 44, 49–64. An, D.J., Roh, I.S., Song, D.S., Park, C.K., Park, B.K., 2007. Phylogenetic characterization of porcine circovirus type 2 in PMWS and PDNS Korean pigs between 1999 and 2006. Virus Research 129, 115–122. Baylis., S.A., Finsterbusch., T., Bannert, N., Blümel, J., Mankertz, A., 2011. Analysis of porcine circovirus type 1 detected in Rotarix vaccine. Vaccine 29, 690–697. Carman, S., Cai, H.Y., DeLay, J., Youssef, S.A., McEwen, B.J., Gagnon, C.A., Tremblay, D., Hazlett, M., Lusis, P., Fairles, J., Alexander, H.S., van Dreumel, T., 2008. The emergence of a new strain of porcine circovirus-2 in Ontario and Quebec swine and its association with severe porcine circovirus associated disease--2004-2006. Canadian Journal of Veterinary Research 72, 259–268. Chae, C., 2005. A review of porcine circovirus 2-associated syndromes and diseases. Veterinary Journal 169, 326–336. Cheung, A.K., 2003. Transcriptional analysis of porcine circovirus type 2. Virology 305, 168–180. Chaiyakul, M., Hsu, K., Dardari, R., Marshall, F., Czub, M., 2010. Cytotoxicity of ORF3 proteins from a nonpathogenic and a pathogenic porcine circovirus. Journal of Virology 84, 11440–11447. Chianini, F., Majó, N., Segalés, J., Domínguez, J., Domingo, M., 2003. Immunohistochemical characterisation of PCV2 associate lesions in lymphoid and non-lymphoid tissues of pigs with natural postweaning multisystemic wasting syndrome (PMWS). Veterinary Immunology and Immunopathology 94, 63-75. Choi, J., Stevenson, G.W., Kiupel, M., Harrach, B., Anothayanontha, L., Kanitz, C.L., Mittal, S. K., 2002. Sequence analysis of old and new strains of porcine circovirus associated with congenital tremors in pigs and their comparison with strains involved with postweaning multisystemic wasting syndrome. Canadian Journal of Veterinary Research 66, 217–224. Cortey M., Olvera A., Grau-Roma L., Segalés J., 2011. Further comments on porcine circovirus type 2 (PCV2) genotype definition and nomenclature. Veterinary Microbiology, 149 522–523. de Castro, A.M., Cortez, A., Heinemann, M.B., Brandão, P.E., Richtzenhain, L.J., 2008. Molecular diversity of Brazilian strains of porcine circovirus type 2 (PCV-2). Research in Veterinary Science 85, 197–200. Darwich, L., Balasch, M., Plana-Durán, J., Segalés, J., Domingo, M., Mateu, E., 2003a. Cytokine profiles of peripheral blood mononuclear cells from pigs with postweaning multisystemic wasting syndrome in response to mitogen, superantigen or recall viral antigens. Journal of Genreal Virology 84, 3453–3457. Darwich, L., Pié, S., Rovira, A., Segalés, J., Domingo, M., 2003b. Cytokine mRNA expression profiles in lymphoid tissues of pigs naturally affected by postweaning multisystemic wasting syndrome. Journal of General Virology 84, 2117–2125. Darwich, L., Segalés, J., Resendes, A., Balasch, M., Plana-Durán, J., Mateu, E., 2008. Transient correlation between viremia levels and IL-10 expression in pigs subclinically infected with porcine circovirus type 2 (PCV2). Research in Veterinary Science 84, 194–198. Dulac, G.C., Afshar, A., 1989. Porcine circovirus antigens in PK-15 cell line (ATCC CCL-33) and evidence of antibodies to circovirus in Canadian pigs. Canadian Journal of Veterinary Research 53, 431–433. Dupont, K., Nielsen, E.O., Baekbo, P., Larsen, L.E., 2008. Genomic analysis of PCV2 isolates from Danish archives and a current PMWS case-control study supports a shift in genotypes with time. Veterinary Microbiology 128, 56–64. Ellis, J., Hassard, L., Clark, E., Harding, J., Allan, G., Willson, P., Strokappe, J., Martin, K., McNeilly, F., Meehan, B., Todd, D., Haines, D., 1998. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome. Canadian Veterinary Journal 39, 44–51. Fan, H., Xiao, S., Tong, T., Wang, S., Xie, L., Jiang, Y., Chen, H., Fang, L., 2008. Immunogenicity of porcine circovirus type 2 capsid protein targeting to different subcellular compartments. Molecular Immunology 45, 653–660. Finsterbusch, T., Mankertz, A., 2009. Porcine circoviruses--small but powerful. Virus Research 143, 177–183. Gagnon, C.A., Tremblay, D., Tijssen, P., Venne, M.H., Houde, A., Elahi, S.M., 2007. The emergence of porcine circovirus 2b genotype (PCV-2b) in swine in Canada. Canadian Veterinary Journal 48, 811–819. Gagnon ,C.A., del Castillo, J.R., Music, N., Fontaine, G., Harel, J., Tremblay, D., 2008. Development and use of a multiplex real-time quantitative polymerase chain reaction assay for detection and differentiation of porcine circovirus-2 genotypes 2a and 2b in an epidemiological survey. Journal of Veterinary Diagnostic Investigation 20, 545–558. Gagnon, C.A., Music, N., Fontaine, G., Tremblay, D., Harel, J., 2010. Emergence of a new type of porcine circovirus in swine (PCV): a type 1 and type 2 PCV recombinant. Veterinary Microbiology 144, 8–23. Gillespie, J., Opriessnig, T., Meng, X.J., Pelzer, K., Buechner-Maxwell, V., 2009. Porcine circovirus type 2 and porcine circovirus-associated disease. Journal of Veterinary Internal Medicine 23, 1151–1163. Grau-Roma, L., Crisci, E., Sibila, M., López-Soria, S., Nofrarias, M., Cortey, M., Fraile, L., Olvera, A., Segalés, J., 2008. A proposal on porcine circovirus type 2 (PCV2) genotype definition and their relation with postweaning multisystemic wasting syndrome (PMWS) occurrence. Veterinary Microbiology 128, 23–35. Hamel, A.L., Lin, L.L., Nayar, G.P., 1998. Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs. Journal of Virology 72, 5262–5267. Hansen, M.S., Pors, S.E., Jensen, H.E., Bille-Hansen, V., Bisgaard, M., Flachs, E.M., Nielsen, O.L., 2010. An investigation of the pathology and pathogens associated with porcine respiratory disease complex in Denmark. Journal of Comparative Pathology 143, 120–131. Harding JC, Ellis JA, McIntosh KA, Krakowka S. Dual heterologous porcine circovirus genogroup 2a/2b infection induces severe disease in germ-free pigs. Veterinary Microbiology 145, 209–219. Hesse, R., Kerrigan, M., Rowland, R.R., 2008. Evidence for recombination between PCV2a and PCV2b in the field. Virus Research 132, 201–207. Horlen, K.P., Schneider, P., Anderson, J., 2007. A cluster of farms experiencing severe porcine circovirus associated disease: Clinical features and association with the PCV2b genotype. Journal of Swine Health Production 15, 270–278. Ihira, M., Yoshikawa, T., Enomoto, Y., Akimoto, S., Ohashi, M., Suga, S., Nishimura N., Ozaki T., Nishiyama Y., Notomi T., Ohta Y., Asano Y., 2004. Rapid diagnosis of human herpesvirus 6 infection by a novel DNA amplification method, loop-mediated isothermal amplification. Journal of Clinical Microbiology 42, 140–145. Juhan, N.M., LeRoith, T., Opriessnig, T., Meng, X.J., 2010. The open reading frame 3 (ORF3) of porcine circovirus type 2 (PCV2) is dispensable for virus infection but evidence of reduced pathogenicity is limited in pigs infected by an ORF3-null PCV2 mutant. Virus Research 147, 60–66. Karuppannan, A.K., Liu, S., Jia, Q., Selvaraj, M., Kwang, J., 2010. Porcine circovirus type 2 ORF3 protein competes with p53 in binding to Pirh2 and mediates the deregulation of p53 homeostasis. Virology 398, 1–11. Kim, J., Chung, H.K., Jung, T., Cho, W.S., Choi, C., Chae, C., 2002. Postweaning multisystemic wasting syndrome of pigs in Korea: prevalence, microscopic lesions and coexisting microorganisms. Journal of Veterinary Medicine Science 64, 57–62. Kim, J., Chae, C., 2005. Necrotising lymphadenitis associated with porcine circovirus type 2 in pigs. Veterinary Record 156, 177–178. Krakowka, S., Hartunian, C., Hamberg, A., Shoup, D., Rings, M., Zhang, Y., Allan, G., Ellis, J.A., 2008. Evaluation of induction of porcine dermatitis and nephropathy syndrome in gnotobiotic pigs with negative results for porcine circovirus type 2. American Journal of Veterinary Research 69, 1615–1622. Lekcharoensuk, P., Morozov, I., Paul, P.S., Thangthumniyom, N., Wajjawalku, W., Meng, X.J., 2004. Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2. Journal of Virolgy 78, 8135–8145 Lefebvre, D.J., Van Doorsselaere, J., Delputte, P.L., Nauwynck, H.J., 2009. Recombination of two porcine circovirus type 2 strains. Archive Virology 154, 875–879. Liu, J., Chen, I., Du, Q., Chua, H., Kwang, J., 2006. The ORF3 protein of porcine circovirus type 2 is involved in viral pathogenesis in vivo. Journal of Virology 80, 5065–5073. Lyoo, K.S., Kim, H.B., Joo, H.S., 2008. Evaluation of a nested polymerase chain reaction assay to differentiate between two genotypes of porcine circovirus-2. Journal of Veterinary Diagnostic Investigation 20, 283–288. Mahé, D., Blanchard, P., Truong, C., Arnauld, C., Le Cann, P., Cariolet, R., Madec, F., Albina, E., Jestin, A., 2000. Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes. Journal of General Virology 81, 1815–1824. Maldonado, J., Segales, J., Martinez-Puig, D., Calsamiglia, M., Riera, P., Domingo, M., Artigas, C., 2005. Identificationof viral pathogens in aborted fetuses and stillborn piglets from cases of swine reproductive failure in Spain. Veterinary Journal 169, 454–456. Meehan, B.M., McNeilly, F., Todd, D., Kennedy, S., Jewhurst, V.A., Ellis, J.A., Hassard, L.E., Clark, E.G., Haines, D.M., Allan, G.M., 1998. Characterization of novel circovirus DNAs associated with wasting syndromes in pigs. Journal of General Virology 79, 2171–2179. Meerts, P., Misinzo, G., McNeilly, F., 2005. Replication kinetics of different porcine circovirus 2 strains in PK-15 cells, fetal cardiomyocytes and macrophages. Archive Virology 150, 427–441. Meng, X.J., 2012. Emerging and Re-emerging Swine Viruses. Transboundary and Emerging Diseases 59, 85–102. Misinzo, G., Delputte P.L., Meerts, P., Nauwynck, H.J., 2006. Porcine circovirus 2 uses heparan sulfate and chondroitin sulfate B glycosaminoglycans as receptors for its attachment to host cells. Journal of Virology 80, 3487–3494. Morozov, I., Sirinarumitr, T., Sorden, S.D., Halbur, P.G., Morgan, M.K., Yoon, K.J., and Paul, P.S., 1998. Detection of a novel strain of porcine circovirus in pigs with postweaning multisystemic wasting syndrome. Journal of Clinical Microbiolgy 36, 2535–2541. Nauwynck, H.J., Sanchez, R., Meerts, P., Lefebvre, D.J., Saha, D., 2012. Cell tropism and entry of porcine circovirus 2. Virus Research 164, 43–45. Nagashima, S., Yoshida, A., Ansai, T., Watari, H., Notomi, T., Maki, K., Takehara, T., 2007. Rapid detection of the cariogenic pathogens Streptococcus mutans and Streptococcus sobrinus using loop-mediated isothermal amplification. Oral Microbiology Immunology 22, 361–368. Niagro, F. D., Forsthoefel, A.N., Lawther, R.P., Kamalanathan, L., Ritchie, B.W., Latimer, K.S., Lukert, P.D., 1998. Beak and feather disease virus and porcine circovirus genomes: intermediates between the geminiviruses and plant circoviruses. Archive Virology 143, 1723–1744. Nieto, D., Aramouni, M., Grau-Roma, L., Segalés, J., Kekarainen, T., 2011. Dynamics of Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2) DNA loads in serum of healthy and postweaning multisystemic wasting syndrome (PMWS) affected pigs. Veterinary Microbiology 152, 284–290. Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N., Hase, T., 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research 28, E63. Olvera, A., Cortey, M., Segalés, J., 2007. Molecular evolution of porcine circovirus type 2 genomes: phylogeny and clonality. Virology 357, 175–185. Opriessnig, T., Thacker, E.L., Yu, S., Fenaux, M., Meng, X.J., Halbur, P.G., 2004. Experimental reproduction of postweaning multisystemic wasting syndrome in pigs by dual infection with Mycoplasma hyopneumoniae and porcine circovirus type 2. Veterinary Pathology 41, 624–640. Opriessnig, T., Janke, B.H., Halbur, P.G., 2006. Cardiovascular lesions in pigs naturally or experimentally infected with porcine circovirus type 2. Journal of Comparative Pathology 134, 105–110. Opriessnig, T., Meng, X.J., Halbur, P.G., 2007. Porcine circovirus type 2 associated disease: update on current terminology, clinical manifestations, pathogenesis, diagnosis, and intervention strategies. Journal of Veterinary Diagnostic Investigation 19, 591–615. Opriessnig, T., Patterson, A.R., Elsener, J., Meng, X.J., Halbur, P.G., 2008. Influence of maternal antibodies on efficacy of porcine circovirus type 2 (PCV2) vaccination to protect pigs from experimental infection with PCV2. Clinical Vaccine Immunology 15, 397–401. Parida, M.M., Santhosh, S.R., Dash, P.K., Tripathi, N.K., Lakshmi, V., Mamidi, N., Shrivastva, A., Gupta, N., Saxena, P., Babu, J.P., Rao, P.V., Morita, K., 2007. Rapid and real-time detection of Chikungunya virus by reverse transcription loop-mediated isothermal amplification assay. Journal of Clinical Microbiology 45, 351–357. Park, J.S., Kim, J., Ha, Y., Jung, K., Choi, C., Lim, J.K., Kim, S.H., Chae, C., 2005. Birth abnormalities in pregnant sows infected intranasally with porcine circovirus 2. Journal of Comparative Pathology 132, 139–144. Pensaert, M.B., Sanchez, R.E. Jr., Ladekjaer-Mikkelsen, A.S., Allan, G.M., Nauwynck, H.J., 2004. Viremia and effect of fetal infection with porcine viruses with special reference to porcine circovirus 2 infection. Veterinary Microbiology 98, 175–183. Phenix, K.V., Weston, J.H., Ypelaar, I., Lavazza, A., Smyth, J.A., Todd, D., Wilcox, G.E., Raidal, S. R., 2001. Nucleotide sequence analysis of a novel circovirus of canaries and its relationship to other members of the genus Circovirus of the family Circoviridae. Journal of General Virology 82, 2805–2809. Quintana, J., Segalés, J., Calsamiglia, M., Domingo, M., 2006. Detection of porcine circovirus type 1 in commercial pig vaccines using polymerase chain reaction. Veterinary Journal 171, 570–573. Saha, D., Lefebvre, D.J., Van Doorsselaere, J., Atanasova, K., Barbé, F., Geldhof, M., Karniychuk, U.U., Nauwynck, H.J., 2010. Pathologic and virologic findings in mid-gestational porcine foetuses after experimental inoculation with PCV2a or PCV2b. Veterinary Microbiology 145, 62–68. Saha, D, Lefebvre, D.J., Ooms, K., Huang, L., Delputte, P.L., Doorsselaere, J.V., Nauwynck, H.J., 2012. Single amino acid mutations in the capsid switch the neutralisation phenotype of porcine circovirus 2. Journal of General Virology 93, 1548–1555. Segalés, J., Allan, G.M., Domingo, M., 2005. Porcine circovirus diseases. Animal Health Research Reviews 6, 119–142. Tischer, I., Mields, W., Wolff, D., Vagt, M., Griem, W., 1986. Studies on epidemiology and pathogenicity of porcine circovirus. Archive Virology 91, 271–276. Tischer, I., Peters, D., Rasch, R., 1987. Replication of porcine circovirus: Induction by glucosamine and cell cycle dependence. Archive Virology 96, 39–57. Todd, D., Weston, J.H., Soike, D., Smyth, J. A., 2001. Genome sequence determinations and analyses of novel circoviruses from goose and pigeon. Virology 286, 354-362. Victoria, J.G., Wang, C., Jones, M.S., Jaing, C., McLoughlin, K., Gardner, S., Delwart, E.L., 2010. Viral nucleic acids in live-attenuated vaccines: detection of minority variants and an adventitious virus. Journal of Virology 84, 6033–6040. Vincent, I.E., Carrasco, C.P., Herrmann, B., Meehan, B.M., Allan, G.M., Summerfield, A., McCullough, K.C., 2003. Dendritic cells harbor infectious porcine circovirus type 2 in the absence of apparent cell modulation or replication of the virus. Journal of Virology 77, 13288–13300. Vincent, I.E., Carrasco, C.P., Guzylack-Piriou, L., Herrmann, B., McNeilly, F., Allan, G.M., Summerfield, A., McCullough, K.C., 2005. Subset-dependent modulation of dendritic cell activity by circovirus type 2. Immunology 115, 388–398. Vincent, I.E., Balmelli, C., Meehan, B., Allan, G., Summerfield, A., McCullough, K.C., 2007. Silencing of natural interferon producing cell activation by porcine circovirus type 2 DNA. Immunology 120, 47–56. Vlasakova, M., Jackova, A., Vilcek, S., 2011. Genetic typing of porcine circovirus type 2 (PCV-2) isolates from Slovakia. Research in Veterinary Science 44, 168–173. Wang, C., Huang, T.S., Huang, C.C., Tu, C., Jong, M.H., Lin, S.Y., Lai, S.S., 2004. Characterization of porcine circovirus type 2 in Taiwan. Journal of Veterinary Medicine Science 66, 469–475. Wang, F., Guo, X., Ge, X.N., Wang, Z.T., Chen, Y.H., Cha, Z.L., Yang, H.C., 2009. Genetic variation analysis of Chinese strains of porcine circovirus type 2. Virus Research 145, 151–156. Yu, S., Halbur, P.G., Thacker, E., 2009. Effect of porcine circovirus type 2 infection and replication on activated porcine peripheral blood mononuclear cells in vitro. Veterinary Immunology and Immunopathogy 127, 350–356. Yu, S., Vincent, A., Opriessnig, T., Carpenter, S., Kitikoon, P., Halbur, P.G., Thacker, E., 2007. Quantification of PCV2 capsid transcript in peripheral blood mononuclear cells (PBMCs) in vitro. Veterinary Microbiology 123, 34–42. Yokota, M., Tatsumi, N., Nathalang, O., Yamada, T., Tsuda, I., 1999. Effects of heparin on polymerase chain reaction for blood white cells. Journal of Clinical Laboratory Analysis 13, 133–140. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16980 | - |
dc.description.abstract | 第二型豬環狀病毒(porcine circovirus type 2, PCV2)為豬隻主要的病毒性病原,其已在世界各地包括台灣的養豬場造成重大的經濟損失。PCV2被認為是引起豬隻離乳後多系統消耗性症候群 (postweaning multisystemic wasting syndrome, PMWS)以及其它相關臨床症狀之主要病原,PCV2所引起的各種臨床症狀被統稱為第二型猪環狀病毒症(PCV2 associated diseases, PCVAD)。PCV2普遍感染於台灣的豬場,但此病毒演化相關資料相當少見;而新穎的分子診斷方法迴路媒介恆溫增幅法(loop-mediated isothermal amplification, LAMP)亦較少使用於此病毒的診斷,包括普遍汙染於PK細胞株的第一型豬環狀病毒(porcine circovirus type 1, PCV1)。因此,本篇論文的目的是進行台灣PCV2病毒的分子流行病學研究,並建立新的臨床診斷技術:迴路媒介恆溫增幅法用以診斷豬環狀病毒(PCV)。在第一項研究中,將2001年至2011年間收集的571株台灣PCV2分離株之基因體全長以聚合酶鏈反應(polymerase chain reaction, PCR)加以增幅並將其產物直接定序。以演化樹分析定序之結果,可發現這些分離株主要分為兩種不同的基因型,PCV2a和PCV2b。其中131株分離株之基因型屬於PCV2a,並可被進一步區分為2B、2D以及2E等3個基因亞型;另外440分離株之基因型屬於PCV2b,並可被進一步區分為1A、1B以及1C等3個基因亞型。 在2001年台灣PCV2分離株主要之基因型為PCV2a,然而自2003年開始,台灣PCV2分離株主要基因型為PCV2b。經由該研究結果分析,台灣PCV2分離株之主要基因型已從PCV2a轉變為PCV2b。在第二項研究中,主要以PCV2第二開放讀碼區(open reading frame 2, ORF2)基因序列為基礎,開發出結合即時監控系統(real-time monitoring system)之LAMP檢測法用以檢測臨床檢體中是否有PCV2a或PCV2b等不同基因型PCV2病毒之單獨或混和感染。LAMP檢測法可以在50分鐘內完成反應,並可同時檢測臨床樣品中PCV2a和PCV2b等不同基因型之分離株,而且並無交叉反應。以LAMP及巢式聚合酶鏈反應(nested PCR)共同檢測含PCV2a和PCV2b基因序列的標準重組質體(recombinant plasmid),LAMP和nested PCR之最低檢出濃度皆為10 copies/μl,顯示LAMP之敏感度等同於nested PCR。將168例臨床檢體同時以LAMP與nested PCR進行檢測,並將nested PCR檢測結果作為金標準(gold standard)以進行比較,LAMP之相對靈敏度(relative sensitivity)為97.7%,相對特異性(relative specificity)為100%。綜合上述研究結果顯示LAMP檢測法可作為一種簡便、快速鑑別PCV2a與PCV2b之診斷工具。在第三項研究中,我們開發出結合real-time monitoring system之LAMP檢測法以檢測PCV1,並應用此法檢測PCV1是否汙染豬用疫苗。LAMP反應可以在一小時內完成,且對於PCV1具有高度特異性,對於其它常見豬隻病毒並不會產生交叉反應。以LAMP及nested PCR共同檢測含PCV1基因序列的recombinant plasmid,LAMP和nested PCR之最低檢出濃度皆為10 copies/μl,顯示LAMP之敏感度等同於nested PCR。另一方面我們收集 25個豬用疫苗並以LAMP和nested-PCR同時進行檢測,其中3個豬用疫苗(11%, 3/25)可同時被LAMP和nested-PCR檢出含PCV1的DNA。LAMP檢測法可作為一個高度特異,敏感,快速診斷技術,並可有效檢測出豬用疫苗是否遭到PCV1 DNA之汙染,為生物製劑之品質做好把關之工作。 | zh_TW |
dc.description.abstract | Porcine circovirus type 2 (PCV2) is one of the major swine viral diseases and caused significant economic loss to pig producers worldwide, including Taiwan. PCV2 has been considered as the causative agent of postweaning multisystemic wasting syndrome (PMWS) as well as other clinical diseases. All these associated syndromes have been categorized as PCV2 associated diseases (PCVAD). PCV2 infection is commonly present in pig farms in Taiwan; however, the information regarding evolution and phylogenic analysis of PCV2 in this area is rare. In addition, application of loop-mediated isothermal amplification (LAMP) in PCV infection, including commonly PK cell line-contaminated PCV1, has rarely been reported. The purposes of this thesis were to perform molecular epidemiological studies of PCV2 in Taiwan and establish a novel diagnostic tool, LAMP, to detect PCV. In the first study, complete genomes of PCV2 were amplified by polymerase chain reaction (PCR) and sequenced directly from 571 Taiwanese PCV2 isolates collected during the period of 2001 and 2011. The phylogenetic tree analysis indicated that these isolates could be divided into 2 distinct genotypes, PCV2a and PCV2b. Among the 571 isolates, 131 isolates were clustered within genotype PCV2a (further classified as 2B, 2D, and 2E), and 440 isolates were clustered within genotype PCV2b (further classified as 1A, 1B, and 1C). PCV2a genotype predominated in 2001, and then PCV2b became the major prevalent genotype since 2003. In the second study, a LAMP method with a real-time monitoring system was developed based on open reading frame 2 (ORF2) in the viral genome for the simultaneous detection and differentiation of PCV2a and PCV2b in clinical samples. The LAMP reaction could be completed within 50 min, and the PCV2a and PCV2b could be detected and accurately distinguished without cross-reaction. The detection limit of the LAMP were 10 copies/μl of the PCV2a and PCV2b recombinant plasmids, demonstrating detection limit comparable to that obtained using nested polymerase chain reaction (nested PCR). On the basis of the results of 168 clinical samples using nested PCR as the gold standard, the relative sensitivity of LAMP was 97.7% and the relative specificity was 100%. Thus, LAMP can be a simple, rapid and valuable tool for the differential diagnosis of PCV2a and PCV2b. In the third study, we have developed a LAMP method with a real-time monitoring system for the detection of PCV type 1 (PCV1) in commercial swine vaccines. This method was highly specific for PCV1, no cross-reaction to other common swine viral pathogens was observed and the LAMP reaction could be completed within one hour. The detection limit of the LAMP for PCV1 DNA was 10 copies/μl of the positive recombinant plasmid, comparable to that obtained from nested PCR. In addition, 25 commercial swine vaccines were tested by both LAMP and nested PCR, and PCV1 DNA was detected in 3/25 commercial swine vaccines (11%) by either method. The LAMP method holds promise for using as a highly specific, sensitive, and rapid diagnostic assay for PCV1 DNA detection in commercial swine vaccines. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T23:51:41Z (GMT). No. of bitstreams: 1 ntu-103-D95629001-1.pdf: 2906907 bytes, checksum: 35de631bf275769e60030eecfdd00b95 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 目錄 Contents
中文摘要 Ⅰ Abstract Ⅲ Chapter I General introduction 1 Chapter II Prevalence and genetic variation of porcine circovirus type 2 in Taiwan from 2001 to 2011 Res. Vet. Sci., 2013, 94, 789-795 25 Chapter III Development and evaluation of a loop-mediated isothermal amplification method for rapid detection and differentiation of two genotypes of porcine circovirus type 2 J. Microbiol. Immuno. Infect., In Press 33 Chapter IV Detection of porcine circovirus type 1 in commercial porcine vaccines by loop-mediated isothermal amplification Folia Microbiol., 2011, 56, 483-489 42 Chapter V General discussion and conclusion 50 | |
dc.language.iso | en | |
dc.title | 臺灣豬環狀病毒分子流行病學之研究 | zh_TW |
dc.title | Molecular Epidemiological Studies of Porcine Circovirus in Taiwan | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-1 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 賴秀穗 | |
dc.contributor.oralexamcommittee | 龐飛,李維誠,張志成,郭村勇 | |
dc.subject.keyword | 豬環狀病毒,迴路媒介恆溫增幅法,樹狀圖分析, | zh_TW |
dc.subject.keyword | Porcine circovirus,loop mediated isothermal amplification,phylogenetic tree analysis, | en |
dc.relation.page | 62 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2014-01-13 | |
dc.contributor.author-college | 獸醫專業學院 | zh_TW |
dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
顯示於系所單位: | 獸醫學系 |
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