【摘要】 本研究旨在比较微芯片电泳及毛细管电泳在急性髓系白血病(AML)FLT3-ITD基因突变检测中的可靠性。应用PCR-微芯片电泳和PCR-毛细管电泳法同时检测214例初治AML患者DNA样本FLT3-ITD基因突变,并通过直接测序法进行序列分析。PCR-微芯片电泳结果显示,FLT3-ITD突变阴性151例,阳性58例(58/214,27.1%),可疑阳性5例(5/214,2.3%);PCR-毛细管电泳结果显示,FLT3-ITD突变阴性147例,阳性67例(67/214,31.3%)。在毛细管电泳结果为阳性的67例样本中,其中4例微芯片电泳结果为阴性,5例为可疑阳性,用直接测序法进行检测,结果均为突变阳性。这一结果揭示,PCR-毛细管电泳法用于检测FLT3-ITD突变更为准确,虽然微芯片电泳组与毛细管电泳组FLT3-ITD突变检测阳性率并无显著差异(卡方检验,P>0.05)。结论:PCR-微芯片电泳与PCR-毛细管电泳均可作为简便的FLT3-ITD基因突变筛查方法,但作为新的检测手段,PCR-毛细管电泳法准确度与测序法一致、可用,而PCR-微芯片电泳法作为诊断检测的方法,其准确度有待于改进。更多还原

【Abstract】 The purpose of the present study was to compare the reliability of microchip electrophoresis and capillary electrophoresis for screening FLT3-ITD gene mutation in acute myeloid leukemia.The FLT3-ITD mutation in the genomic DNA samples from 214 untreated AML patients were separately detected by PCR-microchip electrophoresis and PCR-capillary electrophoresis,then the DNA direct sequencing analysis was carried out.The results from PCR-microchip electrophoresis showed that there were 151 FLT3-ITD mutation negative,58 FLT3-ITD mutation positive(58/214,27.1%) and 5 FLT3-ITD mutation doubtful positive(5/214,2.3%),while the outcomes from PCR-capillary electrophoresis displayed that there were 147 FLT3-ITD mutation negative and 67 FLT3-ITD mutation positive(67/214,31.3%) without doubtful positive.In the 67 FLT3-ITD mutation positive samples detected by using PCR-capillary electrophoresis,4 samples were detected as the negative while 5 samples were measured as the doubtful positive by using PCR-microchip electrophoresis.The follow ed sequencing analysis demonstrated that the above 9 samples were all FLT3-ITD mutation positive,indicating that PCR-capillary electrophoresis was more accurate and sensitive in screening the FLT3-ITD mutation,although statistic analysis showed that there were no significant differences in the detected results between PCR-microchip electrophoresis and PCR-capillary electrophoresis groups(Pearson Chi-squared Test,P >0.05).It is concluded that both PCR-microchip electrophoresis and PCR-capillary electrophoresis were convenient and fast for screening FLT3-ITD mutation,but the accuracy of PCR-microchip electrophoresis awaits further improvement.更多还原