中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2012年
2期
163-166
,共4页
陆滢%汪琼%牧启田%余梦霞%黄勤%金洁
陸瀅%汪瓊%牧啟田%餘夢霞%黃勤%金潔
륙형%왕경%목계전%여몽하%황근%금길
聚丙烯酰胺凝胶电泳%毛细管电泳%急性髓系白血病%FLT3基因%NPM1基因
聚丙烯酰胺凝膠電泳%毛細管電泳%急性髓繫白血病%FLT3基因%NPM1基因
취병희선알응효전영%모세관전영%급성수계백혈병%FLT3기인%NPM1기인
Polyacrylamide gel electrophoresis%Capillary electrophoresis%Acute myeloid leukemia%FLT3 gene%NPM1 gene
目的 建立用聚丙烯酰胺凝胶电泳法(polyacrylamide gel electrophoresis,PAGE)同时快速检测正常核型急性髓细胞白血病(cytogenetically normal acute myeloid leukemia,CN-AML)患者FLT3-ITD 和NPM1基因突变的方法.方法 在多重PCR基础上,用毛细管电泳法(capillary electrophoresis,CE)和PAGE凝胶电泳法同时检测117例初发CN-AML患者的FLT3-ITD和NPM1基因突变.结果 突变型双链DNA分子,如突变型FLT3和NPM1的长度均比野生型长(FLT3 -mut:420 bp＞FLT3-wt:327～332 bp,NPM1 -mut:172 bp＞ NPM1 -wt:168 bp),因此,在PAGE凝胶电泳中突变双链DNA比未突变的DNA移动得慢,从而可将突变检出.117例CN-AML患者均通过CE检测得到验证,其结果与PAGE凝胶电泳结果完全一致(FLT3-ITD +/NPM1-患者18例,占15.4％; FLT3 ITD +/NPM1+患者19例,占16.2％；FLT3-ITD -/NPM1+患者25例,占21.4％；FLT3-ITD -/NPM1-患者55例,占47.0％).结论 两种电泳法均可快速、简便地同时检测CN-AML患者FLT3- ITD和NPM1基因突变.CE检测敏感,图像清晰；PAGE凝胶电泳法则操作简单,成本低,结果可靠,更适于在基层医院开展初步筛查.
目的 建立用聚丙烯酰胺凝膠電泳法(polyacrylamide gel electrophoresis,PAGE)同時快速檢測正常覈型急性髓細胞白血病(cytogenetically normal acute myeloid leukemia,CN-AML)患者FLT3-ITD 和NPM1基因突變的方法.方法 在多重PCR基礎上,用毛細管電泳法(capillary electrophoresis,CE)和PAGE凝膠電泳法同時檢測117例初髮CN-AML患者的FLT3-ITD和NPM1基因突變.結果 突變型雙鏈DNA分子,如突變型FLT3和NPM1的長度均比野生型長(FLT3 -mut:420 bp＞FLT3-wt:327～332 bp,NPM1 -mut:172 bp＞ NPM1 -wt:168 bp),因此,在PAGE凝膠電泳中突變雙鏈DNA比未突變的DNA移動得慢,從而可將突變檢齣.117例CN-AML患者均通過CE檢測得到驗證,其結果與PAGE凝膠電泳結果完全一緻(FLT3-ITD +/NPM1-患者18例,佔15.4％; FLT3 ITD +/NPM1+患者19例,佔16.2％；FLT3-ITD -/NPM1+患者25例,佔21.4％；FLT3-ITD -/NPM1-患者55例,佔47.0％).結論 兩種電泳法均可快速、簡便地同時檢測CN-AML患者FLT3- ITD和NPM1基因突變.CE檢測敏感,圖像清晰；PAGE凝膠電泳法則操作簡單,成本低,結果可靠,更適于在基層醫院開展初步篩查.
목적 건립용취병희선알응효전영법(polyacrylamide gel electrophoresis,PAGE)동시쾌속검측정상핵형급성수세포백혈병(cytogenetically normal acute myeloid leukemia,CN-AML)환자FLT3-ITD 화NPM1기인돌변적방법.방법 재다중PCR기출상,용모세관전영법(capillary electrophoresis,CE)화PAGE응효전영법동시검측117례초발CN-AML환자적FLT3-ITD화NPM1기인돌변.결과 돌변형쌍련DNA분자,여돌변형FLT3화NPM1적장도균비야생형장(FLT3 -mut:420 bp＞FLT3-wt:327～332 bp,NPM1 -mut:172 bp＞ NPM1 -wt:168 bp),인차,재PAGE응효전영중돌변쌍련DNA비미돌변적DNA이동득만,종이가장돌변검출.117례CN-AML환자균통과CE검측득도험증,기결과여PAGE응효전영결과완전일치(FLT3-ITD +/NPM1-환자18례,점15.4％; FLT3 ITD +/NPM1+환자19례,점16.2％；FLT3-ITD -/NPM1+환자25례,점21.4％；FLT3-ITD -/NPM1-환자55례,점47.0％).결론 량충전영법균가쾌속、간편지동시검측CN-AML환자FLT3- ITD화NPM1기인돌변.CE검측민감,도상청석；PAGE응효전영법칙조작간단,성본저,결과가고,경괄우재기층의원개전초보사사.
Objective To establish a stable,rapid multiplex PCR assay combined with PAGE gel electrophoresis for simultaneously detecting FLT3-ITD and NPM1 mutations in acute myeloid leukemia (AML).Methods Capillary electrophoresis (CE) and PAGE gel electrophoresis were simultaneously used to analyze FLT3-ITD and NPM1 mutations in 117 de novo AML patients with normal cytogenetic findings.Results For certain mutations,the length of mutated double-stranded DNA is longer than wild-type DNA.Since FLT3 -mut (420 bp) is longer than FLT3 -wt (327-332 bp),and NPM1 -mut (172 bp) is longer than NPM1-wt (168 bp), heteroduplex will move more slowly during PAGE gel electrophoresis than homoduplex.Therefore the mutations may be detected.A total of 117 CN-AML patients were analyzed with CE and PAGE gel electrophoresis,and the results were identical,which included 18 (15.4％) patients with FLT3-ITD+/NPM1 -,19 (16.2％) patients with FLT3-ITD+/NPM1 +,25 (21.4％) patients with FLT3-ITD-/NPM1+,and 55 (47.0％) patients withFLT3-ITD-/NPM1-.Conclusion Both types of electrophoresis assays may provide a rapid and handy assay for simultaneous detection of FLT3-ITD and NPM1 mutations.CE is relatively sensitive,stable; while PAGE electrophoresis is relatively simple,cheap,and reliable,which may be suitable for primary hospitals and preliminary screening.
