【摘要】 目的探讨在慢性粒细胞白血病(CML)诊断和微小残留病(MRD)监测中,染色体核型分析、荧光原位杂交(FISH)和实时荧光定量PCR(RQ-PCR)的应用价值及其临床意义。方法采用染色体G显带、FISH和RQ-PCR技术对46例CML初发慢性期(CP)患者进行核型分析、BCR/ABL融合基因检测,并对伊马替尼、羟基脲和干扰素治疗后进行追踪观察。结果①46例CML初发患者Ph染色体均为阳性,其中1例伴有附加染色体异常;FISH检测BCR/ABL融合基因均阳性,阳性细胞平均数为(92.20±7.80)%;RQ-PCR检测BCR-ABL/ABL的平均值(68.18±26.67)%。②46例患者随访时间6~24个月,其中17例采用伊马替尼治疗,27例羟基脲治疗,2例干扰素治疗。随访期间,7例出现附加染色体异常,同时FISH检测BCR-ABL融合基因均为阳性(82%~100%),RQ-PCR检测BCR-ABL/ABL值为(51.10±16.30)%,并于0~12个月内进入加速期或急变期。其余39例患者(包括伊马替尼治疗组16例,羟基脲和干扰素治疗组29例),伊马替尼治疗组FISH检测仅4例为阳性(1.0%~5.0%),RQ-PCR检测BCR-ABL/ABL转录本相对平均值(0.10±0.08)%,与治疗前相比明显下降;羟基脲和干扰素治疗组FISH检测均阳性(79%~100%),BCR-ABL/ABL转录本相对平均值(53.65±12.60)%,与治疗前相比无明显改善。结论染色体核型分析、FISH、RQ-PCR能够有效提高CML患者的诊断和治疗后MRD的监测水平。更多还原

【Abstract】 Objective To explore the application and clinical significance of karyotypic analysis,fluorescence in site hybridization(FISH) and real-time quantitative PCR(RQ-PCR) technique for diagnosis and monitoring the minimal residual disease(MRD) in chronic myeloid leukemia(CML).Methods Karyotypes were analyzed by chromosome G banding in 46 CML-CP cases who were monitored the level of BCR-ABL fusion gene by FISH and RQ-PCR before and after administrating imatinib(IM) or hydroxyurea(Hu) or interferon(IFN).Results ①All of 46 cases were detected Ph positive,including one case with extra chromosomal abnormalities.All cases showed positive I-FISH results and the average positive rate was(92.20±7.80)%.The average level of BCR-ABL/ABL transcripts was(68.18±26.67)%.②During the time of follow-up(6-24 months) in 46 cases,BCR-ABL fusion gene was positive(82%-100%) by FISH and the average level of BCR-ABL/ABL transcripts rate was(51.0±16.30)% in 7 cases with extra chromosomal abnormalities,who developed into accelerate phase or blast crisis in the following 0-12 months.The other 39 cases included 16 cases treated with imatinib and 29 cases with Hu or IFN.BCR-ABL fusion gene was positive(1.0%-5.0%) by FISH only in 4 cases treated with imatinib.The average level of BCR-ABL/ABL transcripts was(0.10±0.08)% by RQ-PCR in 16 cases with imatinib,and it was lower than before.BCR-ABL fusion gene was positive(79%-100%) by FISH and the average lwvel of transcripts was(53.65±12.60)% by RQ-PCR in 29 cases treated with hydroxyurea or interferon.Compared to the initial results,there was no improvement.Conclusion It will be greatly improved by karyotypes analysis,FISH and RQ-PCR to diagnose and monitor MRD in CML.更多还原