联合应用FISH和巢式RT-PCR技术检测慢性髓系白血病bcr/abl融合基因

Detection of bcr/abl fusion gene in chronic myeloid leukemia by fluorescence in situ hybridization and nested reverse transcription polymerase chain reaction

目的 检测慢性髓系白血病 (CML)bcr/abl融合基因 ,协助临床诊断。方法 联合应用荧光原位杂交 (FISH)、Northern杂交和巢式RT PCR技术。结果 在 16例被检测的CML患者中 ,有 3例两种方法的结果不一致 ,经Northern杂交验证发现 ,RT PCR较FISH更敏感 ,但存在假阳性 ;另外 ,FISH较容易检测到bcr/abl融合的少见类型 ,而巢式RT PCR需要同时设计多对特殊引物 ,否则极易漏诊。结论 联合应用FISH和巢式RT PCR技术检测CMLbcr/abl融合基因可以优势互补 ,使结果更加准确、可靠 ,更适合于临床诊断、疗效判定以及对微量残留病的监测。

Objective To detect the bcr/abl fusion gene in chronic myeloid leukemia (CML) for assisting in clinical diagnosis. Methods Fluorescence in situ hybridization(FISH) and nested reverse transcription polymerase chain reaction(RT-PCR) were used. Results Out of 16 CML patients, the results of FISH was not consistent with that of nested RT-PCR in 3 cases. When compared with the results of Northern blot, it was found that nested RT-PCR was more sensitive than FISH, but might give false positive results. Moreover, FISH was the easier method for detecting rarer types of bcr/abl transcripts. For detecting the latter occasions nested RT-PCR have to design several specific primers. Conclusion It could take their advantages of applying FISH and nested RT-PCR at the same time to make the results more accurate and reliable for the diagnosis, prognosis and monitor of minimal residual disease.