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研究生:劉素勤
研究生(外文):Su-chen Lou
論文名稱:人類多瘤性病毒,JC病毒,在尿液中排放量與自體免疫疾病關係之研究
論文名稱(外文):Investigation of correlation between the amount of JCV shedding in urine and autoimmune diseases
指導教授:張徳卿
指導教授(外文):D.C.Chang
學位類別:碩士
校院名稱:國立中正大學
系所名稱:分子生物研究所
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:2005
畢業學年度:93
語文別:中文
論文頁數:64
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外文關鍵詞:
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摘要
人類多瘤性病毒,JC病毒,是一種親神經性及致癌性的DNA病毒。人類多瘤性病毒感染人類後可在尿液中偵測到病毒,而尿液中病毒的排放量與飲水相關性是否有關連,以一般而言,飲水多尿液排放也較多,反之亦然。而目前尚未對同人不同時段偵測病毒量相關討論,所以我們用real-time PCR方法偵測JCV病毒同人不同時段病毒的關連性。首先採取大林慈濟醫院住院病人有JCV病毒陽性反應51人每人早上起來第一次尿液與中午時段尿液和晚上時段尿液。我們發現在早上第一次排尿液中,和中午、晚上比較分析,發現早上比中午、晚上病毒量平均較高。此外陽性檢體中,三個時段皆為陽性反應。以此推論早上第一次排放尿偵測病毒量靈敏度最好。目前檢測尿液中病毒量只收集檢體無考慮那一時段收集檢體。此研究結果建議收集固定時段檢體對實驗有更正確的分析與判讀。
張德卿老師實驗室先前的研究發現,在免疫受到抑制的個體中,人類多瘤性病毒出現在尿液中的機率明顯增高。目前推測此病毒感染健康人後,會以潛伏態存在腎臟細胞中,直到宿主免疫受壓抑時才會再活化,進一步可能產生變異或受壓抑基因型再活化。因此,本論文利用同步定量PCR(real-time PCR)的方法來定量尿液中JCV病毒的排放量。我們收集了三類病人的中午時段尿液,第一類為免疫風濕科門診病人(使用低劑量免疫抑制劑量),30人平均年齡44歲。第二類為免疫風濕科住院病人(使用高劑量抑制劑)30人平均年齡48歲。第三類為健檢人(沒有服用免疫抑制劑)20人平均年齡48歲。我們發現使用高劑量免疫抑制劑住院病人的JCV病毒排放量是使用低劑量免疫抑制劑門診病人的5倍。這個結果顯示可能病人服用高劑量抑制劑時,會以潛伏態存在腎臟細胞中,直到免疫受至抑時,使病毒活化複製。
此外,進一步利用Genescan方法分析多型態病毒的基因型,結果發現健檢人與自體免疫疾病門診病人的型態沒有特別差異,住院之自體免疫疾病病人的病毒的多型態基因型變異性較大。我們認為可能因服用高劑量抑制之患者,免疫能力降低,除了一般基因型病毒外,少數會受抑制的基因型病毒而復發,大量繁殖,所以會發現免疫低者,病毒大量製造病毒,以及可能受抑制基因型病毒隨之複製,成為致病性較強的種屬。
Human polyomavirus, JC virus,is a neurotropic and oncogenic DNA virus. Our previous study has proved that the frequency of polyomavirus excretion in urine is apparently increased in immunosuppressed individuals. It has been speculated that the virus may persist in the renal tissue after infecting healthy individuals and be reactivated in immunosuppressive conditions. The virus may also undergo the genetic rearrangement to become epidemic strains. In this study we tried to quantify the shedding of JCV in urine by real-time quantitative PCR. To correctate the amount of JCV shedding in urine and autoimmune diseases.Three groups of subjects were studied. Group I comprised 30 patients (mean age: 44 ) from the Department of Rheumatology at Tzu-Chi Hospital who received low-dose immunosuppressive agents. Group II comprised 20 subjects (mean age: 48 ) selected from physical examination center who did not take immunosuppressive agents. Group III comprised 30 ward patients (mean age: 48 year) who were taking high-dose immunosuppressive agents. We found that the virus shedding ward patients is five times greater than that of the low-dose immunosuppressive agents. patients. The results indicate that the virus may persist be reactivated in renal cells when patients were under high-dose immunosuppressive agent treatment.The virus may be reactivated and replicated until the immune system is restored. In addition, the correlation was also found between JCV shedding and the dose of immunosuppressive agents.These samples were also examined to determine the genotypes of the virus. It was found that there was no difference between healthy individual and OPD patients. More variation of the virval genetypes was found in ward patients.
第一節 前言
一、人類多瘤性病毒(Human polymavirus)簡介……………….1
二、自體免疫疾病(Autoimmune Diseases)簡介……………….1
三、人類多瘤性病毒的分離與基因型的探討分析………………… 2
四、人類多瘤性病毒臨床特徵與診斷……………………………… 5
五、人類多瘤性病毒臨床研究……………………………………… 6
六、人類多瘤性病毒的基因組………………………………………..8
七、本研究的目的與意義…………………………………………….10
第二節 材料與方法…………………………………………………………….13
第三節 結果……………………………………………………………………..22
一、不同時間點尿液中病毒排放量之比較…………………………..22
二、尿液中病毒排放量與至自體免疫疾病之………………………..23
第四節 討論……………………………………………………………………..25
第五節 參考文獻………………………………………………………………..46
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