Cuneo A, Bigoni R, Roberti M G, Bardi A, Balsamo R, Piva N, Castoldi G
Dipartimento di Scienze Biomediche e Terapie Avanzate, Università di Ferrara, Italy.
Haematologica. 1997 Jan-Feb;82(1):85-90.
Over the last 5 years, fluorescence in situ hybridization (FISH) techniques have had an important impact on molecular cytogenetic diagnosis, providing a better understanding of the role of numerical aberrations in hemopoietic neoplasms. The objective of this article is to analyze the clinical applications of FISH in the management of hemopoietic malignancies.
The material examined in the present review includes articles and abstracts published in journals covered by the Science Citation Index and Medline, and personal published and unpublished data.
FISH technology has the advantage of being relatively simple, fast and flexible. Published data and ongoing prospective studies show that, under well-controlled experimental conditions, interphase FISH is more sensitive than conventional metaphase analysis in the detection of numerical abnormalities. Due to the relatively high rate of false positive results, FISH cannot be used for the study of minimal residual disease. However, since molecular strategies for the detection of small-sized aneuploid clones have not been developed yet, FISH represents a useful adjunct to conventional cytogenetics, especially for the quantitation of the size of abnormal clones during the course of the disease and to monitor XX/XY chimerism following sex mis-matched bone marrow transplantation. Different approaches to the study of multiple cell-lineage involvement by chromosome changes have been developed that take advantage of FISH techniques by: a) simultaneous FISH and membrane immunophenotyping of cytologic and histologic preparations; b) two-step analysis based on assessment of the morphology of cells on panoptical stains, with subsequent hybridization and relocation of previously identified cells; c) FISH analysis of enriched cell fractions obtained by cell sorting or by separation of bone marrow cells on a density gradient, and d) study of single hemopoietic colonies grown in semisolid media.
New molecular cytogenetic techniques, such as dual color FISH comparative genomic hybridization, are at hand that will greatly improve the diagnostic power of cytogenetics and make FISH increasingly useful in research laboratories as well as in clinical practice.
在过去5年中，荧光原位杂交（FISH）技术对分子细胞遗传学诊断产生了重要影响，有助于更好地理解造血系统肿瘤中数目异常的作用。本文旨在分析FISH在造血系统恶性肿瘤管理中的临床应用。
本综述所研究的资料包括发表于《科学引文索引》和《医学索引》收录期刊的文章和摘要，以及个人已发表和未发表的数据。
FISH技术具有相对简单、快速且灵活的优点。已发表的数据和正在进行的前瞻性研究表明，在严格控制的实验条件下，间期FISH在检测数目异常方面比传统中期分析更敏感。由于假阳性结果发生率相对较高，FISH不能用于微小残留病的研究。然而，由于尚未开发出检测小尺寸非整倍体克隆的分子策略，FISH是传统细胞遗传学的有用辅助手段，特别是在疾病过程中定量异常克隆的大小以及监测性别不匹配的骨髓移植后的XX/XY嵌合体方面。已经开发出利用FISH技术研究染色体变化累及多个细胞系的不同方法：a）对细胞学和组织学标本同时进行FISH和膜免疫表型分析；b）基于对全景染色细胞形态的评估进行两步分析，随后对先前鉴定的细胞进行杂交和重新定位；c）对通过细胞分选或在密度梯度上分离骨髓细胞获得的富集细胞组分进行FISH分析；d）研究在半固体培养基中生长的单个造血集落。
新的分子细胞遗传学技术，如双色FISH比较基因组杂交，即将出现，这将极大地提高细胞遗传学的诊断能力，并使FISH在研究实验室以及临床实践中越来越有用。
