跳到主要內容

臺灣博碩士論文加值系統

(18.97.14.91) 您好!臺灣時間:2025/02/11 20:04
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:曾淑真
研究生(外文):Su-Cheng Tseng
論文名稱:使用長片段聚合酶連鎖反應與變性高效能液相層析法分析台灣人的血友病基因
論文名稱(外文):Genetic analysis of hemophilia in Taiwanese by using Long-Distance PCR and DHPLC
指導教授:謝豐舟謝豐舟引用關係
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:分子醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:66
中文關鍵詞:長片段聚合酶連鎖反應變性高液相色層分析血友病台灣人
外文關鍵詞:DHPLCTaiwanesehemophiliaLD-PCR
相關次數:
  • 被引用被引用:0
  • 點閱點閱:311
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:1
A型及B型血友病是因為分別缺乏凝血因子VIII和IX所造成的,為最普遍的遺傳性出血性疾病,在人類為性隱性遺傳。其突變型式範圍很廣,包括染色體本身的反轉突變,例如凝血因子VIII的intron 1 和 22,另外還有許多不同型式的突變,包括刪除、插入和點突變所造成的nonsense、 missense和 splicing 位置的突變。血友病病人本身及家屬所遭遇的問題包含了疾病所帶來的痛苦及死亡離別的問題,社會經濟及負擔的問題,防止疾病的再現,帶因者的基因檢測和產前診斷是很重要的。在此篇研究論文中我們結合了次循環長片段聚合酶連鎖反應及變性高液相色層分析法來篩檢大部份的凝血因子VIII和IX的突變。在44個台灣血友病家族中, 29個為A型血友病, 15個為B型血友病,在研究中我們找到了10個突變位置是之前未被發表過的,8個在A型血友病,2個在B型血友病,另外有13個為凝血因子VIII的Intron 22 反轉突變,其餘的突變點之前已描述過。這些突變都經過更進一步的直接定序確認,也沒發現偽陽性或偽陰性。結合次循環長片段聚合酶連鎖分析和變性高效液相色層分析法分析血友病帶因者的基因及產前診斷,是正確性高且快速同時花費較其他傳統方法低的有效方法。
Hemophilia A (HA) and hemophilia B (HB), with the deficiency of coagulation factor VIII (FVIII) and IX (FIX) respectively, represent the most common sex-linked inherited bleeding disorders in human. A wide range of different mutations have been identified including the intrachromosomal inversions involving regions in intron 1 and 22 of the FVIII gene as well as many mutation types found in the remaining part of the factor gene, sunch as large and small deletions, insertions, and point mutations. Patients suffering from those disorders and their families bear great financial and social burden, it is very important to prevent recurrence of the diseases. To achieve this goal genetic analysis for carrier screening and prenatal diagnosis is mandatory. We have established a diagnostic strategy consisting of screening for most common mutations in the Factors VIII and Factor IX genes by using long-distance polymerase chain reaction (LD-PCR)and denaturing high performance liquid chromatography (DHPLC). Forty-four affected Taiwanese families including 29 HA and 15 HB families were analyzed. We found eight novel point mutations in 6 HA families and 2 HB families together with two novel small deletion mutations in HA family.The intron 22 inverions in 13 HA families and the remaining mutations have been described previously in HA and HB mutation database. These small mutations were further confirmed by direct sequencing. Neither false positive nor false negative results were found. Our combinatory approach by subcycling LD-PCR and heteroduplex analysis based on DHPLC proves to be a highly informative, rapid and practical means to detect mutations in affected individuals and carriers of hemophilias.
簡稱或縮寫對照表…………………………………………………4
中英文摘要………………………………………………………..6
一、 前言…………………………………………………………8
二、 緒論…………………………………………………………11
血友病的歷史沿革…………………………………………12
臨床表現……………………………………………………13
遺傳表現……………………………………………………14
篩檢試驗……………………………………………………14
分子遺傳試驗………………………………………………14
產前檢查……………………………………………………15
遺傳諮詢…………………………………………………..15
處理…………………………………………………………16
三、 研究動機……………………………………………………17
四、 實驗材料與儀器……………………………………………18
五、 實驗方法及步驟……………………………………………20
六、 結果…………………………………………………………23
七、 討論…………………………………………………………25
附録一. DHPLC的分析原理………………………………………….31
附錄二. 次循環長片段聚合酶連鎖反應之原理…………………34
附錄三. A型血友病國外報導突變型式所占的比例………………35
附錄四. B型血友病國外報導突變型式所占的比例……………36
附錄五. 凝血因子VIII的結構圖………………………………37
圖表……………………………………………………………….38
參考資料………………………………………………………….63
Bagnall RD, Waseem N, Green PM, Giannelli F, 2002. Recurrent inversion breaking intron 1 of the factor VIII gene is a frequent cause of severe hemophilia A. Blood 99:168-74.
Castaldo G, Nardiello P, Bellitti F, Santamaria R, Rocino A, Coppola A, di Minno G, Salvatore F, 2003. Haemophilia B: from molecular diagnosis to gene therapy. Clin Chem Lab Med 41:445-51.
Drayna D, White R, 1985. The genetic linkage map of the human X chromosome. Science 230:753-8.
Erfle V, Hehlmann R, Mellert W, Kruger G, Seifried E, Heimpel H, Rasokat H, Lechler E, Holzer E, Hellstern P, et al., 1985. Prevalence of antibodies to HTLV-III in AIDS risk groups in West Germany. Cancer Res 45:4627s-4629s.
Francis RB, Jr., Kasper CK, 1983. Reproduction in hemophilia. Jama 250:3192-5.
Gitschier J, Wood WI, Goralka TM, Wion KL, Chen EY, Eaton DH, Vehar GA, Capon DJ, Lawn RM, 1984. Characterization of the human factor VIII gene. Nature 312:326-30.
Goodeve AC, 1998. Advances in carrier detection in haemophilia. Haemophilia 4:358-64.
Huber CG, Oefner PJ, Bonn GK, 1993. High-resolution liquid chromatography of oligonucleotides on nonporous alkylated styrene-divinylbenzene copolymers. Anal Biochem 212:351-8.
Ingerslev J, Schwartz M, Lamm LU, Kruse TA, Bukh A, Stenbjerg S, 1989. Female haemophilia A in a family with seeming extreme bidirectional lyonization tendency: abnormal premature X-chromosome inactivation? Clin Genet 35:41-8.
Ivaskevicius V, Jurgutis R, Rost S, Muller A, Schmitt C, Wulff K, Herrmann FH, Muller CR, Schwaab R, Oldenburg J, 2001. Lithuanian haemophilia A and B registry comprising phenotypic and genotypic data. Br J Haematol 112:1062-70.
Jones AC, Austin J, Hansen N, Hoogendoorn B, Oefner PJ, Cheadle JP, O''Donovan MC, 1999. Optimal temperature selection for mutation detection by denaturing HPLC and comparison to single-stranded conformation polymorphism and heteroduplex analysis. Clin Chem 45:1133-40.
Kemball-Cook G, Tuddenham EG, Wacey AI, 1998. The factor VIII Structure and Mutation Resource Site: HAMSTeRS version 4. Nucleic Acids Res 26:216-9.
Kraus EM, Brettler DB, 1988. Assessment of reproductive risks and intentions by mothers of children with hemophilia. Am J Med Genet 31:259-67.
Laken SJ, Petersen GM, Gruber SB, Oddoux C, Ostrer H, Giardiello FM, Hamilton SR, Hampel H, Markowitz A, Klimstra D, Jhanwar S, Winawer S, Offit K, Luce MC, Kinzler KW, Vogelstein B, 1997. Familial colorectal cancer in Ashkenazim due to a hypermutable tract in APC. Nat Genet 17:79-83.
Lakich D, Kazazian HH, Jr., Antonarakis SE, Gitschier J, 1993. Inversions disrupting the factor VIII gene are a common cause of severe haemophilia A. Nat Genet 5:236-41.
Leuer M, Oldenburg J, Lavergne JM, Ludwig M, Fregin A, Eigel A, Ljung R, Goodeve A, Peake I, Olek K, 2001. Somatic mosaicism in hemophilia A: a fairly common event. Am J Hum Genet 69:75-87.
Liu Q, Nozari G, Sommer SS, 1998. Single-tube polymerase chain reaction for rapid diagnosis of the inversion hotspot of mutation in hemophilia A. Blood 92:1458-9.
Liu Q, Sommer SS, 1998. Subcycling-PCR for multiplex long-distance amplification of regions with high and low GC content: application to the inversion hotspot in the factor VIII gene. Biotechniques 25:1022-8.
Ljung R, Tedgard U, 2003. Genetic counseling of hemophilia carriers. Semin Thromb Hemost 29:31-6.
Ljung RC, Sjorin E, 1999. Origin of mutation in sporadic cases of haemophilia A. Br J Haematol 106:870-4.
O''Donovan MC, Oefner PJ, Roberts SC, Austin J, Hoogendoorn B, Guy C, Speight G, Upadhyaya M, Sommer SS, McGuffin P, 1998. Blind analysis of denaturing high-performance liquid chromatography as a tool for mutation detection. Genomics 52:44-9.
Peake I, 1998. Genetic services available for counselling and prenatal diagnosis of haemophilia. Haemophilia 4 Suppl 2:24-5.
Pittman DD, Alderman EM, Tomkinson KN, Wang JH, Giles AR, Kaufman RJ, 1993. Biochemical, immunological, and in vivo functional characterization of B-domain-deleted factor VIII. Blood 81:2925-35.
Rosendaal FR, Smit C, Briet E, 1991. Hemophilia treatment in historical perspective: a review of medical and social developments. Ann Hematol 62:5-15.
Rosner F, 1969. Hemophilia in the Talmud and rabbinic writings. Ann Intern Med 70:833-7.
Saenko EL, Ananyeva N, Kouiavskaia D, Schwinn H, Josic D, Shima M, Hauser CA, Pipe S, 2002. Molecular defects in coagulation Factor VIII and their impact on Factor VIII function. Vox Sang 83:89-96.
Schwaab R, Brackmann HH, Meyer C, Seehafer J, Kirchgesser M, Haack A, Olek K, Tuddenham EG, Oldenburg J, 1995. Haemophilia A: mutation type determines risk of inhibitor formation. Thromb Haemost 74:1402-6.
Sumita DR, Vainzof M, Campiotto S, Cerqueira AM, Canovas M, Otto PA, Passos-Bueno MR, Zatz M, 1998. Absence of correlation between skewed X inactivation in blood and serum creatine-kinase levels in Duchenne/Becker female carriers. Am J Med Genet 80:356-61.
Thompson AR, 2003. Structure and function of the factor VIII gene and protein. Semin Thromb Hemost 29:11-22.
Tuddenham EG, Cooper DN, Gitschier J, Higuchi M, Hoyer LW, Yoshioka A, Peake IR, Schwaab R, Olek K, Kazazian HH, et al., 1991. Haemophilia A: database of nucleotide substitutions, deletions, insertions and rearrangements of the factor VIII gene. Nucleic Acids Res 19:4821-33.
Varekamp I, Suurmeijer T, Brocker-Vriends A, Rosendaal FR, 1992. Hemophilia and the use of genetic counseling and carrier testing within family networks. Birth Defects Orig Artic Ser 28:139-48.
Varekamp I, Suurmeijer TP, Brocker-Vriends AH, van Dijck H, Smit C, Rosendaal FR, Briet E, 1990. Carrier testing and prenatal diagnosis for hemophilia: experiences and attitudes of 549 potential and obligate carriers. Am J Med Genet 37:147-54.
Varekamp I, Suurmeijer TP, Rosendaal FR, Brocker-Vriends AH, 1993. The use of preventive health care services: carrier testing for the genetic disorder haemophilia. Soc Sci Med 37:639-48.
Wollina K, Bowen DJ, Syrbe G, Zintl F, 1993. Female twins with severe Christmas disease (hemophilia B). Thromb Haemost 70:774-6.
Xie YG, Zheng H, Leggo J, Scully MF, Lillicrap D, 2002. A founder factor VIII mutation, valine 2016 to alanine, in a population with an extraordinarily high prevalence of mild hemophilia A. Thromb Haemost 87:178-9.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top