Michalová K
3rd Medical Department, General Faculty Hospital, Prague, Czech Republic.
Folia Biol (Praha). 1996;42(6):311-4.
The principle of the fluorescence in situ hybridization (FISH) method is in the base pairing of the DNA probe to complementary sequences in the studied specimen. The hybridization of specific DNA or RNA probes to the cellular targets attached to the microscopic slides is widely used for the identification of chromosomal translocations, deletions, amplifications of specific genes, and chromosome number changes in mitotic and/or interphase cells. The use of FISH with the modifications of the basic method meant a breakthrough in detection and diagnosis of human malignancies. During the last tow years FISH was used in our laboratory for: (a) identification of constitutive and acquired numerical and structural chromosomal abnormalities; (b) detection of minimal residual disease or early relapse in patients treated for leukemia by bone marrow transplantation (BMT) and/or chemotherapy; (c) determination of the cytogenetic pattern of non-dividing or terminally differentiated cells. To confirm the structural rearrangements found by the classical G-banding technique, the whole chromosome painting probes which hybridize to multiple chromosomal sequences were used. The alpha-satellite DNA probes which detect centromeric repetitive sequences were utilized for determining the numerical and sex chromosome changes. Specific unique chromosomal sequences which can confirm all chromosomal rearrangements, i.e., deletions, translocations or inversions with the corresponding breakpoints were introduced for specific cases. Recently, every chromosomal translocation, deletion and any other structural or numerical change found by conventional cytogenetic analysis in the bone marrow cells of the patients with leukemia has been verified in our laboratory by FISH. The results of this study showed that FISH is more efficient than conventional cytogenetics in detecting residual malignant cells. For chromosomal rearrangements FISH is an extremely sensitive method which not only verifies but also interprets with more precision the findings of classical cytogenetics.
荧光原位杂交（FISH）方法的原理是DNA探针与所研究标本中的互补序列进行碱基配对。特定DNA或RNA探针与附着在显微镜载玻片上的细胞靶标的杂交被广泛用于识别染色体易位、缺失、特定基因的扩增以及有丝分裂和/或间期细胞中的染色体数目变化。对基本方法进行改进后的FISH应用意味着人类恶性肿瘤检测和诊断方面的一项突破。在过去两年中，FISH在我们实验室用于：（a）识别组成性和获得性染色体数目和结构异常；（b）检测接受骨髓移植（BMT）和/或化疗的白血病患者的微小残留病或早期复发；（c）确定非分裂或终末分化细胞的细胞遗传学模式。为了确认经典G带技术发现的结构重排，使用了与多个染色体序列杂交的全染色体涂染探针。检测着丝粒重复序列的α卫星DNA探针用于确定染色体数目和性染色体变化。针对特定情况引入了能够确认所有染色体重排（即具有相应断点的缺失、易位或倒位）的特定独特染色体序列。最近，我们实验室通过FISH验证了白血病患者骨髓细胞中通过传统细胞遗传学分析发现的每一种染色体易位、缺失以及任何其他结构或数目变化。这项研究的结果表明，FISH在检测残留恶性细胞方面比传统细胞遗传学更有效。对于染色体重排，FISH是一种极其灵敏的方法，它不仅能验证经典细胞遗传学的发现，还能更精确地解释这些发现。
