In Situ-RT PCR定量检测16例慢性粒细胞白血病仇bcr/abl融合基因

Detection of bcr/abl Fusion Transcript in Chronic Myeloid Leukemia by in Situ Reverse Transcription- Polymerase Chain Reaction

目的 研究原位逆转录多聚酶链反应技术（Ｉｎ Ｓｉｔｕ－ＲＴ ＰＣＲ）在检测慢性粒细胞白血病（ＣＭＬ）ｂｃｒ／ａｂｌ融合基因中的应用价值及其意义。方法：用 Ｉｎ Ｓｉｔｕ－ＲＴ ＰＣＲ方法测定 ＣＭＬ病人ｂｃｒ／ａｂｌ融合基因表达量。结果：在不破坏细胞完整性条件下细胞系Ｋ５６２和ＣＭＬ病人检测到ｂｃｒ／ａｂｌＲＮＡ的扩增产物，表现为细胞浆内出现蓝紫色颗粒，计算这种阳性细胞的百分率来相对定量地检测ＣＭＬ病人ｂｃｒ／ａｂｌ ｍＲＮＡ的表达水平，这种检测技术的敏感性达１０－４水平以上。结论：Ｉｎ Ｓｉｔｕ－ＲＴ ＰＣＲ可以相对定量地检测ＣＭＬ病人ｂｃｒ／ａｂｌ ｍＲＮＡ的表达，直观地评价α－干扰素（α－ＩＦＮ）、骨髓移植（ＢＭＴ）等治疗ＣＭＬ的疗效，本检测方法敏感性高，又可用做微小残留病（ＭＲＤ）的检测。

Objective: To evaluate the application and significance of in situ reverse transcription-polymerase chain reaction (in situ-RT PCR) in the detection of the bcr/abl fusion-transcript in patients with chronic myeloid leukemia(CML). Methods:In situ-RT PCR was used to detect the expression of bcr/abl fusion transcript. Results: Without the destruction of the cells, the amplification products of the bcr/abl mRNA in the K562 cel1 line and CML patients were detected, displaying the distinct blue cytoplasmic granules in the positive cells. The expression level of the bcr/abl mRNA was expressed as the percentage of the positive cells. The sensitivity of this method was over l0-4. Conclusion: In Situ-RT PCR can be used to quantitively detect the relative expression level of the bcr/abl mRNA in CML patients and to valuate the effectiveness of α- interferon[hu2](α-IFN)or bone marrow transplantation (BMT) in the treatment of CML. It can also be used in the detection of the minimal residual disease(MRD)of CML because of its high sensitivity.