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首页> 外文期刊>Mutation Research, C. Mutation Research Letters >ADVANTAGES AND LIMITATIONS OF USING FLUORESCENCE IN SITU HYBRIDIZATION FOR THE DETECTION OF ANEUPLOIDY IN INTERPHASE HUMAN CELLS
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ADVANTAGES AND LIMITATIONS OF USING FLUORESCENCE IN SITU HYBRIDIZATION FOR THE DETECTION OF ANEUPLOIDY IN INTERPHASE HUMAN CELLS

机译：荧光原位杂交检测间质人细胞中非融合的优势与局限性

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摘要

Fluorescence in situ hybridization with chromosome-specific DNA probes is being increasingly utilized for the detection of chromosome aberrations induced in vitro and in vivo by chemical and physical agents. Although potentially a powerful technique, FISH studies for aneuploidy can be heavily influenced by cellular phenomena and hybridization artifacts which make the performance and interpretation of the results difficult. As a consequence, frequently hyperdiploid frequencies are reported in the literature which are substantially higher than one would expect based upon frequencies seen in conventional metaphase analyses. In this article, a number of the potential pitfalls that we have encountered while performing FISH analyses for aneuploidy are discussed and their potential impact on the observed hybridization frequencies is described. After considering these factors, the frequencies of lymphocyte nuclei containing 3 and 4 chromosome copies are compared between metaphase values obtained from published human population studies and interphase values obtained from similar studies using FISH. It is concluded that by using caution in the evaluation of slides, interphase studies using FISH to detect hyperdiploidy and polyploidy can provide estimates of numerical alterations which closely reflect those seen during metaphase analysis using either FISH or conventional approaches. However, due to the inability of interphase analysis to distinguish hyperdiploidy from polyploidy as well as other potential problems, frequencies of aneuploid nuclei obtained using single label FISH should only be considered approximations of absolute frequencies. For additional accuracy, multi-color FISH with two or more different probes should be performed.
机译：与染色体特异性DNA探针的荧光原位杂交技术正越来越多地用于检测化学和物理试剂在体外和体内诱导的染色体畸变。尽管可能是一种强大的技术，但非整倍性的FISH研究可能会受到细胞现象和杂交伪像的严重影响，这使得其性能和结果解释变得困难。结果，在文献中报道了频繁的超二倍体频率，该频率大大高于基于常规中期分析中看到的频率所期望的频率。在本文中，我们讨论了在进行非整倍性FISH分析时遇到的许多潜在陷阱，并描述了它们对观察到的杂交频率的潜在影响。在考虑了这些因素之后，将包含3和4个染色体拷贝的淋巴细胞核的频率在从已发表的人群研究获得的中期值与从使用FISH的相似研究获得的中期值之间进行比较。结论是，通过谨慎地评估载玻片，使用FISH检测超二倍体和多倍体的相间研究可以提供数值变化的估计值，这些变化紧密反映了使用FISH或常规方法进行中期分析时看到的变化。但是，由于无法进行相间分析来区分多倍体和多倍体以及其他潜在问题，使用单标签FISH获得的非整倍体核的频率应仅视为绝对频率的近似值。为了获得更高的准确性，应使用带有两个或多个不同探针的多色FISH。

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