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| 題 名 | 檢測骨髓增生性疾病之JAK2V617F點突變應以特異性對偶基因聚合酶連鎖反應法適時配合基因定序才能兼顧特異性及敏感度=Maximize Sensitivity and Specificity of JAK2V617F Detection in Myeloproliferative Disorder by Combined Use of Allele-specific Polymerase Chain Reaction and DNA Sequencing |
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| 作 者 | 許瑋眞; 林哲斌; 吳柏燕; 趙淑貞; 陳如珍; 李龍雄; | 書刊名 | Journal of Biomedical & Laboratory Sciences |
| 卷 期 | 22:3 2010.09[民99.09] |
| 頁 次 | 頁95-101 |
| 分類號 | 415.12 |
| 關鍵詞 | 基因定序; 特異性對偶基因聚合酶連鎖反應; JAK2V617F點突變; 骨髓增生性疾病; Sequencing; Allele-specific polymerase chain reaction; AS-PCR; JAK2V617F point mutation; Myeloproliferative disorder; MPD; |
| 語 文 | 中文(Chinese) |
| DOI | 10.30046/JBLS.201009.0003 |
| 中文摘要 | JAK2V617F點突變在骨髓增生性疾病(myeloproliferative disorder, MPD)具有臨床診斷的價值,本文 探討是否可參照國內外文獻報導以敏感度高、耗時較短的特異性對偶基因聚合酶連鎖反應(allelespecific polymerase chain reaction,AS-PCR)方法直接檢測這種單點突變。本研究利用AS-PCR、 基因定序(sequencing)檢驗36例實驗組及36例對照組之JAK2V617F點突變。以AS-PCR法檢測陽性結 果與基因定序結果一致性達100%,以AS-PCR法檢測陰性結果卻出現一例MPD檢體在PCR引子結 合序列發生第二個點突變導致的偽陰性結果(dbSNP C/T allele,mRNA pos=2342/Protein pos= 616(Huref:NM004972/NP004963)),使AS-PCR特異性降為基因定序的98.6%。因此檢測JAK2V617F 點突變時建議先以根據文獻報導敏感度較基因定序高的AS-PCR檢測,當結果為陽性時可確定為 JAK2V617F點突變,當結果為陰性時一定要再加做基因定序避免偽陰性結果,這樣應用於臨床實驗 室才能兼顧特異性及敏感度。 |
| 英文摘要 | Objective: JAK2V617F mutation has been used for clinical diagnosis in myeloproliferative disorders (MPD). This study investigated the use of an efficient and highly sensitive method, allele-specific polymerase chain reaction (AS-PCR), for the detection of this single point mutation. In this study, we determined the sensitivity and specificity of AS-PCR for calling JAK2V617F point mutation by comparing the results of 36 experimental and 36 control samples to those derived by sequencing as the gold standard. All AS-PCR positive samples were also positive by sequencing. However, there was one (2.8%) false negative AS-PCR call of MPD sample due to second mutation in the PCR primer binding sequence(dbSNP C/T allele,mRNA pos=2342/Protein pos=616(Huref:NM004972/ NP004963)), resulting in a specificity of 98.6%. Based on these results, we recommend a two-step strategy to test JAK2V617F mutation first by the more efficient AS-PCR method, and only re-test negative samples by gene sequencing to maintain both specificity and sensitivity while improving the efficiency in clinical laboratories. |
本系統中英文摘要資訊取自各篇刊載內容。