Multiplex ligation-dependent probe amplification for the detection of chromosomal gains and losses in formalin-fixed tissue
Multiplex ligation-dependent probe amplification for the detection of chromosomal gains and losses in formalin-fixed tissue
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DOI：
10.1097/01.pas.0000146701.98954.47
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发表时间：
2005-03-01
影响因子：
--
通讯作者：
Jeuken, JW
中科院分区：
文献类型：
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作者：
van Dijk, MC;Rombout, PD;Jeuken, JW
福尔马林固定石蜡包埋组织的分子分析在组织病理学诊断和肿瘤研究中具有重要意义。多重连接依赖性探针扩增（MLPA）是一种可用于在一个实验中检测多达45个不同DNA序列的拷贝数改变的技术。该技术可以部分降解DNA，这使得该技术非常适合于分析福尔马林固定的病变。我们通过分析从福尔马林固定的黑色素瘤中分离的DNA来测试MLPA的可靠性，所述黑色素瘤先前通过比较基因组杂交（CGH）来表征，并且另外通过分析29个常规处理的黑色素细胞病变来测试MLPA的适用性。MLPA似乎是一种可靠和有效的方法来评估DNA拷贝数的变化，86%的基因座测试显示一致的CGH结果。不一致主要涉及MLPA检测到而CGH未检测到的变化，可能是由于CGH分辨率较低和偶尔的假阳性MLPA结果的组合。对于MLPA在诊断环境中的应用，应在特定的感兴趣区域使用不同的探针，以防止假阳性MLPA结果。在研究和诊断环境中，MLPA是一种快速筛查大量福尔马林固定病变DNA获得和丢失的技术。
Molecular analysis on formalin-fixed paraffin-ernbedded tissue is of increasing importance in diagnostic histopathology and tumor research. Multiplex ligation-dependent probe amplification (MLPA) is a technique that can be used for detection of copy number alterations of up to 45 different DNA sequences in one experiment. It call be performed oil partially degraded DNA, which makes this technique very suitable for analysis of formalin-fixed lesions. We tested the reliability of MLPA by analyzing DNA isolated from formalin-fixed melanomas that were previously characterized by comparative genomic hybridization (CGH), and additionally the applicability of MLPA was tested by analyzing 29 routinely processed melanocytic lesions. MLPA appears to be a reliable and efficient method to evaluate DNA copy number changes as 86% of the loci tested revealed concordant CGH results. Discordance mainly involved alterations that were detected by MLPA and not by CGH probably due to a combination of lower resolution of CGH and occasionally false positive MLPA results. For application of MLPA in a diagnostic setting, different probes on a specific region of interest should be used to prevent false positive MLPA results. In a research setting as well as in a diagnostic setting, MLPA is a fast technique to screen large numbers of formalin-fixed lesions for DNA gains and losses.