多重荧光原位杂交结合染色体涂抹技术检测骨髓增生异常综合征复杂核型异常

Analysis of complex chromosomal aberrations in patients with myelodysplastic syndromes using multiplex fluorescence in situ hybridization combined with whole chromosome painting

目的探讨多重荧光原位杂交（multiplex fluorescence in situ hybridization,M-FISH）及全染色体涂抹（whole chromosome painting,WCP）技术在骨髓增生异常综合征（myelodysplastic syndromes,MDS）复杂核型异常检测中的价值。方法对7例常规R显带具有复杂染色体异常的MDS患者应用M-FISH技术确定复杂染色体的重排及标记染色体的组成,识别微小易位。并进一步采用双色WCP技术验证M-FISH检测的结果。结果M-FISH不仅证实了R显带的结果,而且确定了R带核型分析没有确定的6种标记染色体、9种有不明来源的额外物质增加的染色体、5种衍生染色体的组成和来源及4种被忽略的微小易位。涉及17号染色体的异常及-5/5q-是MDS最为常见的两种染色体异常。WCP技术纠正了一些M-FISH漏检及误检的异常。结论M-FISH是明确复杂染色体异常的很有用的分子生物学工具,WCP是M-FISH技术的重要补充,R带核型分析结合分子细胞遗传学工具M-FISH和WCP可以更加准确地描述复杂染色体异常。

Objective To explore the value of multiplex fluorescence in situ hybridization （M-FISH） in combination with whole chromosome painting （WCP） in the detection of complex chromosomal aberrations （CCAs） in myelodysplastic syndromes （MDS）. Methods M-FISH was used in seven MDS patients with R-banding CCAs to refine the complex chromosomal rearrangements, and to identify cryptic translocations and characterization of marker chromosomes. Dual-color WCP procedures were further performed in 7 cases to confirm some rearrangements detected by M- FISH. Results M-FISH confirmed all results of R-banding. The composition and origin of 6 kinds of marker chromosomes, 9 kinds of chromosomes with additional material undetermined and 5 kinds of derivative chromosomes undefined by conventional cytogenetics （CC） were defined after M-FISH analysis; four kinds of cryptic translocations overlooked by CC were found on derivative chromosomes and previously normal appearing chromosomes. In addition, M-FISH revealed some nonrandom aberrations： aberrations involving chromosome 17 and - 5/5q - were the two most frequent aberrations. Some misclassified and missed chromosomal aberrations by M-FISH were corrected by WCP. Conclusion MFISH is a powerful molecular cytogenetic tool in clarification of CCAs. Complementary WCP helps us to identify misclassifted and missed chromosomal aberrations by M-FISH. CC in combination with molecular cytogenetic techniques, such as M-FISH and WCP, can unravel complex chromosomal aberrations more precisely.