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題名 | Quantitative Analysis of Chimerism after Allogeneic Peripheral Blood Stem Cell Transplantation=異體周邊血液幹細胞移植後骨髓植入之定量分析 |
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作者 | 陳定平; 曹國倩; 王博南; 曾慶平; 孫建峰; | 書刊名 | 長庚醫學 |
卷期 | 25:11 2002.11[民91.11] |
頁次 | 頁734-742 |
分類號 | 416.175 |
關鍵詞 | 周邊血液幹細胞移植; 定量短斷片重複序列基因分析法; 基因嵌合體; Peripheral blood stem cell transplantation; PSCT; Quantitative short tandem repeats; STRs; Mixed chimerism; |
語文 | 英文(English) |
中文摘要 | 背景: 針對異體周邊血液幹細胞移植,目前許多植入評估的分析方法,包括限制酵素片段長度多型性 (restriction fragment length polymorphism) 分析法及多型性基因座 (polymorphic genetic loci) 的聚合酵素連鎖反應(polymerase chain reaction) 分析法,已逐漸取代傳統的評估法。為了增進周邊血液幹細胞植入評估方法的效能,本實驗室建立一套可定量的非放射性短片段重複序列基因(quantitative short tandem repeat; quantitative STR) 分析法。 方法: 移植前先將受髓者及捐贈者的周邊白血球DNA萃取出來,再利用包含九個STR的檢驗套組 (AmpFISTR Profiler Plus Kit) 將受髓者及捐贈者的STR複製擴大,經過基因定序儀 (ABI PRISM 377 DNA sequencer) 及GeneScan 2.1分析軟體處理後,我們可找出有效的基因等位點 (informative alleles) 用來區分受贈者及捐贈者間不同處。為了定量分析,我們必須另外以不同比例混合受贈者及捐贈者的DNA以製作標準曲線,接者就能以此標準曲線客觀評估受贈者有多少比例的基因復發。 結果: 本篇研究共回溯收集十位幹細胞移植病患,其中兩位受髓者在移植後追蹤的檢體發現有基因嵌合體 (chimerism) 產生,顯示可能白血病復發。一位在第三個月追蹤時,發現38.7%的DNA復發;另一位在第十個月追蹤發現有6.5%的DNA復發。 結論: 這套方法可提供正確、敏感、且可定量及早期偵測復發的移植後追蹤分析,以提供臨床醫師更多的資訊,協助其採取適當的醫療行為。 |
英文摘要 | Background: For peripheral blood stem cell transplantation (PSCT), several engraftment analysis methods have been performed including detection of restriction fragment length polymorphisms and amplification of polymorphic genetic loci. To facilitate monitoring of the engraftment, a quantitative, non-isotopic method using a short tandem repeat (STR) maker has been set up in our laboratory. Methods: DNAs from pretransplant recipients and donors were amplified with the AnpFISTR Profiler Plus kit that contains 9 STR markers. The fluorescent polymerase chain reaction products were then fractionated on polyacrylamide gels in an ABI PRISM 377 DNA sequencer. Results were analyzed using GeneScan 2.1 software. We selected the best markers as informative alleles which can distinguish donor from recipient. For quantitative analysis of the engraftment, we prepared a mixed chimeric sample by mixing pretransplant recipient and donor DNAs in different ratios to produce a standard curve. After amplifying the posttransplant recipient DNA, we were able to detect the extent of engraftment by interpolating the percent peak area of the informative alleles from this standard curve. Result: We retrospectively analyzed 10 patients who had received allogeneic PSCT. Two of them showed some degree of mixed chimerism indicating leukemic relapse. In case one, 38.7% of the recipient DNA was first detected in the third month after PSCT. In case two, 6.5% of the recipient DNA was first detected in the tenth month after PSCT. Conclusion: In summary, this method provides an accurate, quantitative, and early assessment of mixed chimerism in posttransplant patients. Such information may be useful to guide implementation of additional treatment to circumvent graft failure or replase in the future. |
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