流式免疫微球捕获技术和荧光定量PCR检测BCR-ABL融合基因的方法学比较

摘要

Objective To evaluate cytometric bead assay method by comparing it with real-time quantitative PCR method in detection of BCR-ABL fusion gene. Methods 18 chronic myeloid leukemia(CML) patients,6 acute myeloblastic leukemia(AML) patients,9 iron deficiency anemia(IDA) patients were selected. BCR-ABL mRNA and protein in their bone marrow were detected by real-time quantitative PCR method and cytometric bead assay method separately.RSSUIl8 11le results from 6 AML cases and 9 IDA caseswere all negative.The real-time quantitative PCR result showed that there were 2 cases ofPl90 type and 16 cases ofP210 type.At thesame time,the samples were all positive detecting by cytometric bead assay method.A correlation coefficient of lgMFI detected by cyto-metric bead assay and lgCopies detected by real-time quantitative PCR WaS 0.708(P<0.01).Conclusion Compared to real一timequantitative PCRmethod,cytometric bead assay method has a hish sensitivity for different transcripts,but it is unable to indentify typesof transcript.Cytometric bead assay in detection of BCR.ABL is suitalde for initial screening detection.%目的:对流式微球捕获技术和荧光定量PCR技术在BCR-ABL融合基因中的检测进行比较和评价.方法:选取18例慢性粒细胞性白血病(chronic myeloid leukemia,CML)患者、6例急性髓系白血病(acute myeloblastic leukemia,AML)患者、9例缺铁性贫血(iron deficiency anemia,IDA)患者,分别用流式微球捕获技术和荧光定量PCR技术检测其骨髓BCR-ABL融合基因蛋白和mRNA.结果:6例AML和9例IDA患者均未检出阳性.18例CML患者荧光定量PCR检出16例P210和2例P190,流式微球捕获技术则均检出阳性.流式微球捕获技术的1gMFI(平均荧光强度)和荧光定量PCR法的lgCopies(拷贝数)的相关系数为0.708(P ＜0.01).结论:流式免疫微球捕获技术对BCR-ABL各种断裂类型敏感性高,但不能区分断裂类型,适合作为初筛实验.