癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2014年
2期
123-127
,共5页
李爽%陆雪%赵骅%封江彬%刘青杰
李爽%陸雪%趙驊%封江彬%劉青傑
리상%륙설%조화%봉강빈%류청걸
泛着丝粒探针%荧光原位杂交%染色体畸变%简并引物PCR
汎著絲粒探針%熒光原位雜交%染色體畸變%簡併引物PCR
범착사립탐침%형광원위잡교%염색체기변%간병인물PCR
pan-centromeric probe%FISH%chromosome aberrations%degenerate oligonucleotide priming-PCR
目的：建立一种检测辐射诱导染色体畸变的快速荧光原位杂交(FISH)方法。方法：两步简并引物PCR法扩增人α-卫60星DNA，制备荧光基团直接标记的泛着丝粒探针；对 Coγ射线照射后的人外周血淋巴细胞染色体标本进行FISH分析，荧光显微镜下检测着丝粒及染色体形态。结果：直接标记泛着丝粒探针杂交后显示所有染色体着丝粒均有较强的信号；应用制备的FISH探针检测到γ射线照射诱导的双着丝粒、着丝粒环和易位染色体畸变，37℃杂交5-12 h后经过简单的洗涤即可在荧光显微镜下检测到较好的信号。结论：本研究制备的直接标记泛着丝粒探针，可用于FISH快速检测辐射诱导的染色体双着丝粒体、着丝粒环和易位畸变。目的：建立一种检测辐射诱导染色体畸变的快速荧光原位杂交(FISH)方法。方法：两步简并引物PCR法扩增人α-卫60星DNA，制备荧光基团直接标记的泛着丝粒探针；对 Coγ射线照射后的人外周血淋巴细胞染色体标本进行FISH分析，荧光显微镜下检测着丝粒及染色体形态。结果：直接标记泛着丝粒探针杂交后显示所有染色体着丝粒均有较强的信号；应用制备的FISH探针检测到γ射线照射诱导的双着丝粒、着丝粒环和易位染色体畸变，37℃杂交5-12 h后经过简单的洗涤即可在荧光显微镜下检测到较好的信号。结论：本研究制备的直接标记泛着丝粒探针，可用于FISH快速检测辐射诱导的染色体双着丝粒体、着丝粒环和易位畸变。
目的：建立一種檢測輻射誘導染色體畸變的快速熒光原位雜交(FISH)方法。方法：兩步簡併引物PCR法擴增人α-衛60星DNA，製備熒光基糰直接標記的汎著絲粒探針；對 Coγ射線照射後的人外週血淋巴細胞染色體標本進行FISH分析，熒光顯微鏡下檢測著絲粒及染色體形態。結果：直接標記汎著絲粒探針雜交後顯示所有染色體著絲粒均有較彊的信號；應用製備的FISH探針檢測到γ射線照射誘導的雙著絲粒、著絲粒環和易位染色體畸變，37℃雜交5-12 h後經過簡單的洗滌即可在熒光顯微鏡下檢測到較好的信號。結論：本研究製備的直接標記汎著絲粒探針，可用于FISH快速檢測輻射誘導的染色體雙著絲粒體、著絲粒環和易位畸變。目的：建立一種檢測輻射誘導染色體畸變的快速熒光原位雜交(FISH)方法。方法：兩步簡併引物PCR法擴增人α-衛60星DNA，製備熒光基糰直接標記的汎著絲粒探針；對 Coγ射線照射後的人外週血淋巴細胞染色體標本進行FISH分析，熒光顯微鏡下檢測著絲粒及染色體形態。結果：直接標記汎著絲粒探針雜交後顯示所有染色體著絲粒均有較彊的信號；應用製備的FISH探針檢測到γ射線照射誘導的雙著絲粒、著絲粒環和易位染色體畸變，37℃雜交5-12 h後經過簡單的洗滌即可在熒光顯微鏡下檢測到較好的信號。結論：本研究製備的直接標記汎著絲粒探針，可用于FISH快速檢測輻射誘導的染色體雙著絲粒體、著絲粒環和易位畸變。
목적：건립일충검측복사유도염색체기변적쾌속형광원위잡교(FISH)방법。방법：량보간병인물PCR법확증인α-위60성DNA，제비형광기단직접표기적범착사립탐침；대 Coγ사선조사후적인외주혈림파세포염색체표본진행FISH분석，형광현미경하검측착사립급염색체형태。결과：직접표기범착사립탐침잡교후현시소유염색체착사립균유교강적신호；응용제비적FISH탐침검측도γ사선조사유도적쌍착사립、착사립배화역위염색체기변，37℃잡교5-12 h후경과간단적세조즉가재형광현미경하검측도교호적신호。결론：본연구제비적직접표기범착사립탐침，가용우FISH쾌속검측복사유도적염색체쌍착사립체、착사립배화역위기변。목적：건립일충검측복사유도염색체기변적쾌속형광원위잡교(FISH)방법。방법：량보간병인물PCR법확증인α-위60성DNA，제비형광기단직접표기적범착사립탐침；대 Coγ사선조사후적인외주혈림파세포염색체표본진행FISH분석，형광현미경하검측착사립급염색체형태。결과：직접표기범착사립탐침잡교후현시소유염색체착사립균유교강적신호；응용제비적FISH탐침검측도γ사선조사유도적쌍착사립、착사립배화역위염색체기변，37℃잡교5-12 h후경과간단적세조즉가재형광현미경하검측도교호적신호。결론：본연구제비적직접표기범착사립탐침，가용우FISH쾌속검측복사유도적염색체쌍착사립체、착사립배화역위기변。
OBJECTIVE: To establish a rapid fluorescence in situ hybridization (FISH) method and to analyze the chromosome aberrations induced by irradiation.METHODS:Human centromeric alpha satellite DNA was amplified and directly labeled with Cy3-dUTP or Fluorescein-12-dUTP by degenerate oligonucleotide priming-PCR (DOP-PCR) to prepare two pan-centromeric probes. Chromosome aberrations in lymphocytes of healthy irradiated peripheral blood 60samples with Coγ-rays were analyzed with FISH.RESULTS:Two directly labeled pan-centromeric probes hybridized to the centromeres of all 46 chromosomes. The established rapid FISH method could easily detect chromosome aberrations after hybridizing 5 h to 12 h in 37 ℃ and simple washing.CONCLUSION: FISH method established in this study could be use to rapidly detect the dicentrics,centric rings,and translocations induced by irradiation.OBJECTIVE:To establish a rapid fluorescence in situ hybridization (FISH) method and to analyze the chromosome aberrations induced by irradiation. METHODS:Human centromeric alpha satellite DNA was amplified and directly labeled with Cy3-dUTP or Fluorescein-12-dUTP by degenerate oligonucleotide priming-PCR (DOP-PCR) to prepare two pan-centromeric probes. Chromosome aberrations in lymphocytes of healthy irradiated peripheral blood 60 samples with Coγ-rays were analyzed with FISH. RESULTS:Two directly labeled pan-centromeric probes hybridized to the centromeres of all 46 chromosomes. The established rapid FISH method could easily detect chromosome aberrations after hybridizing 5 h to 12 h in 37℃and simple washing. CONCLUSION:FISH method established in this study could be use to rapidly detect the dicentrics,centric rings,and translocations induced by irradiation.
