骨髓染色体联合FISH技术在恶性血液病诊断中的应用价值  

Application value of bone marrow chromosome combined with fluorescence in situ hybridization technique in the diagnosis of hematological malignancies

作　　者：李付广 张晓静 谢小雷 汤素环 唐江 吴爱娟 尹卫国 LI Fuguang;ZHANG Xiaojing;XIE Xiaolei;TANG Suhuan;TANG Jiang;WU Aijuan;YIN Weiguo(Molecular Diagnostic Center,The Sixth Affiliated Hospital of Guangzhou Medical University,Qingyuan People’s Hospital,Qingyuan Guangdong 511518,China;Department of Pharmacy,The Sixth Affiliated Hospital of Guangzhou Medical University,Qingyuan People’s Hospital,Qingyuan Guangdong 511518,China)

机构地区：[1]清远市人民医院分子诊断中心·广州医科大学附属第六医院,广东清远511518 [2]清远市人民医院药学部·广州医科大学附属第六医院,广东清远511518

出　　处：《中国实验诊断学》2024年第3期292-297,共6页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金青年项目(81803523);清远市科技计划项目(2019A036,2022KJJH036)。

摘　　要：目的探究联合改良骨髓染色体制备方法和荧光原位杂交(FISH)技术,分析其在恶性血液病特别是在慢性粒细胞白血病(CML)中的临床应用价值。方法收集2019年1月至2022年6月来清远市人民医院就诊的355例恶性血液病患者作为研究对象,首先抽取16例患者的骨髓细胞对骨髓染色体制备的接种浓度、秋水仙碱浓度、时间和低渗等进行改良,优化后对培养时间进行单因素方差分析,按照不同培育时间(16 h、20 h、24 h和28 h)将每例平均分成A、B、C和D 4组;最后用改良方法对所有患者进行骨髓染色体核型分析,同时利用FISH技术对CML患者进行BCR-ABL融合基因检测,并对异常染色体统计分析。结果单因素方差分析发现4组染色体分裂相均值差异有统计学意义(P<0.05),且B组(20 h)显著高于D组(P<0.05);B组的培养成功率同样显著高于D组(P<0.05)。355例恶性血液病患者均一性培养成功率为96.6%(343/355),异常染色体检出率为40.3%(143/355)。CML患者骨髓染色体和FISH阳性检出率一致,在加速期和急变期检出额外异常染色体。结论改良后的骨髓染色体短期培养法成功率高、稳定性好、异常染色体检出率高,培养在16~24 h之间为宜,20 h最为理想,不宜超过28 h;Ph染色体和额外异常染色体与CML病程密切相关,异常染色体核型的检出为临床对白血病的诊断分型、后续治疗及预后判断提供了重要参考依据。Objective To evaluate the clinical application of improved and optimized method for preparing bone marrow chromosomes combined with fluorescence in situ hybridization(FISH)technique in the diagnosis of hematological malignancies,especially in chronic myeloid leukemia(CML).Methods A total of 355patients with hematological malignancies in Qingyuan People’s Hospital from January 2019to June 2022were enrolled in this study.Firstly,marrow cells from 16patients were employed to optimized the bone marrow chromosomes preparing method.Then,each case was equally divided into 4groups(A,B,C and D groups)according to the different culture time(16,20,24and 28h)and one-way analysis of variance was performed.The improved method was then used to detect bone marrow karyotype in all patients.In addition,FISH technique was used to detect the BCR-ABL fusion gene in CML patients,and abnormal chromosomes were statistically analyzed.Results One-way analysis of variance found that there was a significant difference in the mean value of chromosome division among the four groups(P<0.05),and group B(20h)was significantly higher than group D(28h,P<0.05).The culture success rate of group B was also significantly higher than that of group D(P<0.05).The uniform culture success rate of 355patients with hematological malignancies was 96.9%(344/355),and the detection rate of abnormal chromosomes was 40.3%(143/355).The positive detection rates of chromosomes and FISH in the bone marrow of CML patients were consistent,and additional abnormal chromosomes were detected in the accelerated phase and blast phase.Conclusion The improved short-term culture method with 20 hours of culture has high success rate,good stability,and high detection rate of abnormal chromosomes,which is the best solution for the preparation of bone marrow chromosomes.Ph chromosomes and additional abnormal chromosomes are closely related to the course of CML.The detection of chromosome abnormalities provides an important reference for clinical diagnosis,classification,subsequent

关 键 词：骨髓染色体 荧光原位杂交 异常染色体核型 恶性血液病 

分 类 号：R730.263[医药卫生—肿瘤]