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研究生:林等義
研究生(外文):Teng-Yi
論文名稱:利用毛細管電泳探討JC人類多瘤性病毒多形性與臨床應用
論文名稱(外文):Polymorphism Studies of Polyomavirus JC and Its Clinical Application using Capillary Electrophoresis Analysis
指導教授:蔡淦仁
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2006
畢業學年度:95
語文別:中文
論文頁數:54
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Polyomavirus JC(JCV)病毒基因型分析,傳統方式是利用限制酶切割片段多型性分型及核苷酸定序方式來偵測與分析,這些基因型分析主要是以病毒的地理演化特性來分類,然而為探討這些病毒的基因型對於致病性和診斷有密切的關係,所以本研究進一步應用毛細管電泳技術,測試過去已知在臺灣流行的JCV基因型,包括CY、TW-1、TW-2、TW-3等,基因體中的調節區域VP1(Regulatory region of VP1),核苷酸序列約400 bp長度間的變異性。本研究首先利用PCR偵測尿液檢體中的JCV,發現無服用免疫抑制劑的健康者50人中有10人為陽性(20%),腎臟移植60人中有24人(40%)呈陽性反應,再將陽性反應的尿液檢體利用毛細管電泳技術偵測基因型,結果顯示10人JCV陽性無服用免疫抑制劑的健康者中有8人(80%)僅有單一基因型,2人(20%)則有二種基因型存在。而在腎臟移植病患24人JCV呈陽性反應的尿液中可偵測到5人(21%)僅有單一基因型,16人(66%)有二種基因型,3人(12%)具有三種以上基因型病毒存在,這與過去以限制酶切割片段多型性分型的研究報告,只能偵測到一種基因型的存在有明顯的差異。同時本研究所使用DNA複製的產物量只需要傳統方法PCR的1/200即可偵測,顯示毛細管電泳分析比傳統方法具有更高的靈敏度,將有助於臨床診斷及追蹤移植病患,因為服用免疫抑制劑可能產生JCV病毒變異的評估,以及病毒變異與所引起併發症嚴重度的相關性。同時我們相信這種方法未來也可以應用在其他容易產生抗藥性變異株的病毒例如BKV、HBV、HCV、HIV等之研究。

The genotyping of polyomaviruses JC virus (JCV) has been traditionally determined by restriction fragment length polymorphism analysis and by nucleotide sequencing. These genotypes are primarily associated with geographically based virus evolution. To investigate its implication in the pathogenesis and diagnosis of JCV infection, this study aims to further assess the genetic polymorphism in the regulatory region of VP1 of CY, TW-1, TW-2 and TW-3 genotypes which predominate in Taiwanese.
A 400-bp amplified fragment of the regulatory region of VP1 was detected in 20% of urine samples from healthy adults receiving no immunosuppressants (n = 50). Twenty-four out of sixty (40%) kidney transplantation patients on immunosuppression medication were found positive for the VP1 amplification of JCV DNA. The genotypes of these amplified DNA fragments were further analyzed by capillary electrophoresis. It was found that 80% (n = 10) of healthy adults carried one single genotype whereas the rest carried two genotypes. 21% and 66% of JCV-positive kidney transplantation patients (n =24) respectively carried one and two genotypes whereas the remaining showed multi-genotype. Our result is in sharp contrast to previous findings by restriction fragment length polymorphism analysis which only detected one single genotype. It is worth noting that the amount of DNA used by this approach is 0.5% less than that by PCR analysis, suggesting greater sensitivity of capillary electrophoresis analysis. Such a technique can thus provide a new insight into the clinical diagnosis of JCV infection. Moreover, it will facilitate our ability to develop an effective therapeutic strategy for complications associated with immunosuppressant-induced JCV mutation in kidney transplantation patients. It is conceivable that the technique can be further implemented in the study of drug-resistance mutation of viruses such as BKV, HBV, HCV, HIV, etc.


目錄
壹、中英文摘要......................1
貳、縮寫檢索表......................5
參、前言.........................6
肆、實驗動機.......................17
伍、實驗材料與方法....................18
陸、結果.........................24
柒、討論.........................26
捌、參考文獻....................... 30
玖、圖表與圖表說明....................39


1. Agostini HT, Ryschkewitsch CF, and Stoner GL.(1996) Genotype profile of human polyomavirus JC excreted in urine of immunocompetent individuals. J. Clin. Microbiol. 34:159-164.
2. Aksamit AJ, Sever JL, Major EO.(1986) Progressive multifocal leukoencephalopathy: JC virus detection by in situ hybridization compared with immunohistochemistry. Neurology 36: 499-504.
3. Apperley JF, Rice SJ, Bishop JA, Chia YC, Krausz T, Gardner SD, and Goldmam JM.(1987) Late-onset hemorrhagic cystitis associated with urinary excretion of polyomaviruses after bone marrow transplantation. Transplantation 43:108-112.
4. Arthur RR, Shah KV, Baust SJ, Santos GW, and Saral R.(1986) Association of BK viruria with hemorrhagic cystitis in recipients of bone marrow transplants. N. Engl. J. Med. 315:230-234.
5. Chang DC, Tsai RT, Wang ML, and Ou WC.(1996) Different genotypes of human polyomaviruses found in patients with autoimmune diseases in Taiwan. J. Med. Virol. 48:204-209.
6. Chang DC, Wang ML, Ou WC, Lee MS, Ho HN, and Tsai RT.(1996) Genotype of human polyomaviruses in urine sample of pregnant individuals in Taiwan. J. Med. Virol. 48:95-101
7. Coleman DV, Mackenzie EFD, Gardner SD, Poulding JM, Amer B, and Russell WJI.(1978) Human polyomavirus(BK) infection and ureteric stenosis in renal allograft recipients. J. Clin. Pathol. 31:338-347.
8. Coleman DV, Wolfendale MR, Daniel RA, Dhanial NK, Gardner SD, Gibson PE, and Field AM.(1980) A prospective study of human polyomavirus infection in pregnancy. J. Infect. Dis. 142:1-8.
9. Daniel R, Shah K, Madden D, and Stagno S.(1981) Serological Investigation of the possibility of congenital transmission of papovavirus JC. Inf. Imm. 33:319-321.
10. Chang D, Sugimoto C, Wang M, Tsai RT, and Yogo Y.(1999) JC virus genotypes in Taiwan aboriginal tribe(Bunun): implications for its population history. Arch Virol 144:1081-1090
11. DiMail D, and Nathans D.(1980) Cold sensitive regulatory mutants of simian virus 40. J. Mol. Virol. 140: 192-142
12. Dorries K, Vogel E, Gunther S, and Czub S.(1994) Infection of human polyomaviruses JC and BK in peripheral blood leukocytes from immunocompetent individuals. Virol. 198:59-70.
13. Dorries K, Loeber G, and Meixensberger J.(1987) Association of polyomaviruses JC, SV 40 and BK with human brain tumors. Virology 160,268-270.
14. Dubois V, Dutronc H, and Lafon ME.(1997) Latency and reactivation of JC virus in peripheral blood of human immunodeficiency virus type Ι-infected patients. J. Clin. Microbiol. 35:2288-2292.
15. Frisque RJ, Bream GL, and Cannella MT.(1984) Human polyomavirus JC virus genome. J. Virol. 51:458-469.
16. Gardner SD, MacKenzie EF, Smith C, and Porter AA.(1984) Prospective study of the human polyomaviruses BK and JC and cytomegalovirus in renal transplant recipients. J. Clin.Pathol.37:578-586
17. Gerard R, and Gluzman Y.(1986) Functional analysis of the role the AT-rich region and upstream flanking sequences in simian virus 40 DNA replication. Mol. Cell Biol. 6:4570-4577.
18. Gibson PE, Field AM, Gardner SD, and Coleman DV.(1981) Occurrence of IgM antibodies against BK and JC polymaviruses during pregnancy. J. Clin. Pathol. 34:674-679.
19. Horgan TF, Padgett GL, Walker DL, Borden EC, and Frias ZJL.(1983) Survey of human polyomavirus (JCV, BKV) infections in 139 patients with lung cancer, breast cancer, melanoma, or lymphoma. In: Madden DL, Liss AR, eds. Polyomaviruses and Human Neurological Disease, Severe.New York : 311–24.
20. Horgan TF, Borden EC, McBain HA, Padgett GL, and Walker DL.(1980) Human polyomavirus infection with JC virus and BK virus in renal transplant patients. Ann, Intern. Med. 92:373-378
21. Iida T, Kitamura T, Guo J, Taguchi F, Aso Y, Nagashima K. and Yogo Y.(1993) Origin of JC polyomavirus variants associated with progressive multifocal leukoencephalopathy. Proc.Natl. Acad. Sci.USA. 90:5062-5065.
22. John RP, Anthony OO, Amin M, Deborah AP.(2003) Capillary electrophoresis and its application in the clinical laboratory. Clinica Chimica Acta 330:1-30
23. Kasuen W, Anima S, Soumya A, Elizabeth FM, Lowery R, Carine VL, Daniel JS, Dennis KB, and Robert H.(2005) HIV-1 Tat Interactions with p300 and PCAF Transcriptional Coactivators Inhibit Histone Acetylation and Neurotrophin Signaling through CREB. J. Bio. Chem. Vol.280, No.10:9390-9399
24. Khalili K, and White MK.(2006) Human demyelinating disease and the polyomavirus JCV. Multiple Sclerosis 12:133-142
25. Kitamura T, Aso Y, Kuniyoshi N, Hara K. and Yogo Y.(1990) High incidence of urinary JC virus excretion in nonimmunosuppressed older patients. J. Inf. Dis. 161, 1128-1133.
26. Kitamura T, Kunitake T, Guo J, Tominaga T, Kawabe K, and Yogo Y.(1994) Transmission of the human polyomavirus JC virus occurs both within the family and outside the family. J. Clin. Microbiol. 32,2359-2363.
27. Kumar KU, Pater A, and Pater MM.(1993) Human JC virus perfect palindromic nuclear factor 1-binding sequences important for glial cell-specific expression in differentiating embryonal carcinoma cells. J Virol. Jan;67(1):572–576
28. Ledoux S, Libman I, Robert F, et al:(1989) Progressive multifocal leukoencephalopathy with gray matter involvement. Can J Neurol Sci 16; 200-202.
29. Lee MH, Chen TZ, Wang LS, Yen PS, Hsu YH.(2007) Progressive Multifocal Leukoencephalopathy in an AIDS Patient. J Formos. Assoc. Mar.; 106(3 Suppl):S24-28
30. Liu SQ, Tung CH, Huang KY, Lin LH, Lai NS.(2006) Quantitative Measurement of Urinary JC virus Shedding by Real-Time PCR at Different Time Intervals – a Useful Guide for Clinical and Laboratory References. Tzu Chi Med J.18.No.3
31. Loeber G, and Dorries K.(1988) DNA rearrangements in organ-specific variants of polyomavirus JC strain GS. J. Virol. 62:1730-1735.
32. Major EO, Amemiya K, Tornatore CS, et al:(1992) Pathogenesis and molecular biology of progressive multifocal leukoencephalopathy, the JC virus-induced demyelinating disease of the human brain. Clin Microbiol Rev 5: 49-73
33. Markowitz RB, Eaton BA, Kubik MF, Latorra D, Mcgregor JA, and Dynan WS.(1991) BK virus and JC virus shed during pregnancy have predominantly archetypal regulatory regions. J. Virol. 65,4515-4519.
34. Martin JD, and Foster GC.(1984) Multiple JC virus genomes from one patient. J.Gen.Virol.65,1405-1411.
35. Munoz-Marmol AM, Mola G, Fernandez-Vasalo A, et al:(2004) JC virus early protein detection by immunohistochemistry in progressive multifocal leukoencephalopathy: A comparative study with in situ hybridization and polymerase chain reaction. J Neuropathol Exp Neurol. Nov;63(11):1124-30.
36. Padgett BL, Rogers CM, and Walker DL.(1997) JC Virus, a Human Polyomavirus Associated with Progressive Multifocal Leukoencephalopathy: Additional Biological Characteristics and Antigenic Relationships. Infect. Immun. 15:356-366.
37. Peterin BM, and Trono D.(2003) Hide, shield and strike back: How HIV-infected cells avoid immune eradication. Nature reviews/Immunology Vol. 3:97-107
38. Riccardo DS, Alberta A.(2000) Use of Denaturing gradient gel electrophoresis for human polyomavirus JC sequence analysis. J. Virol. Methods. 85;101-108
39. Sabath BF, Major EO.(2002) Traffic of JC virus from sites of initial infection to the brain: the path to progressive multifocal leukoencephalopathy. J Infect Dis.186 (Suppl 2): 180-186.
40. Silverman L, and Rubinstein LL.(1965) Electron microscopic observations in a case of progressive multifocal leukoencephalopathy. Acta Neuropathol. 5: 215-224
41. Sugimoto C, Kitamura T, Guo J, Mohammed NA, Shchelkunov SN, Otova B, Paul O, Chollet JY, Sayda ES, Mohamed E, Gerard G, Kocagoz T, Sanong C, Kyaw ZT, Thein S, Moe K, Kobayashi N, Taguchi F, and Yogo Y.(1997) Typing of urinary JC virus DNA offers a novel means of tracing human migrations. Proc. Natl. Acad. Sci.USA Vol. 94, pp.9191-9196, August

42. Tada H, and Khalil K.(1992) A novel sequence-specific DNA-binding protein, LCP-1, interacts with single-stranded DNA and differentially regulates early gene expression of the human neurotropic JC virus. J.Virol. 66,6885-6892.
43. Taguchi F, Kajioka J, and Miyamura T.(1982) Prevalence rate and age of acquisition of antibodies against JC and BK virus in human sera. Microbio. Imm. 26:1057-1064
44. Tooze J.(1981) DNA tumor viruses, 2nd ed. Cold spring Harbor. NY: Cold spring Harbor Laboratory. 61370.
45. Tsai RT, Wang ML, Ou WC, Lee YL, Li SY, Fung CY, Huang YL, Tzeng TY, Chang DC.(1997) Incidence of JC virus is higher than that of BK virus in Taiwan. J. Med. Virol. 52:253-257.
46. Walker DL.(1978) Progressive multifocal leukoencephalopahy: An opportunistic viral infection of the central nrvous system. In Vinken PJ, Bryun GW (eds): Handbook of Clinical Neurology. Amsterdam, North-Holland Publishing Co.18:307-329.
47. Wang ML, Tzeng TY, Fung CY, Ou WC, Tsai RT, Chen CH, and Chang DC.(1999) Human anti-JC virus serum reacts with native but not denatured JC major capsid protein VP1. J. Virol. Methods. 78:171-176.
48. Wang ML, Tsai RT, Ou WC, Lin CK, Tsay GJ, Chang H, and Chang DC.(2000) Treatment with cytotoxic immunosuppression agents increases urinary excretion of JCV in patients with autoimmune disease. J.Med.Virol. 62:505-510.
49. Willoughby E, Price RW, Padgett BL, Walker DL, and Dupont B.(1980) Progressive multifocal leukoencephalopathy (PML): in vitro cell-mediated immune responses to mitogens and JC virus. Neurology. 30:256-262.
50. Yogo Y, Iida T, Taguchi F, Kitamura T, and Aso Y.(1991) Typing of Human Polyomavirus JC Virus on The Basis of Restriction Fragment Length Polymorphisms. J. Clin. Microbiol. OCT., p. 2130-2138.
51. ZuRhein GM, and Chou SM.(1968) Papovavirus in progressive multifocal leukoencephalopathy. Res. Publ. Assoc. Nerve. Ment. Dis. 44:307


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