Natté Remco, van Eijk Ronald, Eilers Paul, Cleton-Jansen Anne-Marie, Oosting Jan, Kouwenhove Mathilde, Kros Johan M, van Duinen Sjoerd
Department of Pathology, Leiden University Medical Center, P.O. box 9600, 2300RC, Leiden, The Netherlands.
Brain Pathol. 2005 Jul;15(3):192-7. doi: 10.1111/j.1750-3639.2005.tb00520.x.
Multiplex ligation-dependent probe amplification (MLPA) is a new assay for the detection of multiple chromosomal deletions in tumor tissue in a single experiment. Since genotyping of gliomas with oligodendroglial features by the detection of 1p/19q chromosomal deletions became essential for treatment decisions, we developed and validated an MLPA-based assay to determine these losses in formalin fixed and paraffin embedded oligodendroglial tumors (OG). Nineteen OGs, and 10 control samples were analyzed by MLPA and the results were correlated with those obtained by fluorescent in situ hybridization (FISH). The MLPA results were reproducible in all samples in which repeated experiments were performed. In 18 of 19 OGs, MLPA and FISH were concordant for presence or absence of 1p deletion. In 3 OGs, MLPA detected a 19q deletion not shown by FISH. For the other 15 OGs, MLPA and FISH were concordant. In one sample with 50% to 75% of tumor, MLPA failed to detect the 1p/19q deletions revealed by FISH (though with borderline values of significance). We conclude that MLPA is a valid and reproducible method for the detection of 1p/19q chromosomal deletions in OGs stored on formalin fixed, paraffin embedded tissue.
多重连接依赖探针扩增(MLPA)是一种可在单次实验中检测肿瘤组织中多个染色体缺失的新检测方法。由于通过检测1p/19q染色体缺失对具有少突胶质细胞特征的胶质瘤进行基因分型对于治疗决策至关重要,我们开发并验证了一种基于MLPA的检测方法,以确定福尔马林固定石蜡包埋的少突胶质细胞瘤(OG)中的这些缺失情况。通过MLPA对19个OG和10个对照样本进行分析,并将结果与荧光原位杂交(FISH)获得的结果进行关联。在所有进行重复实验的样本中,MLPA结果均可重复。在19个OG中的18个中,MLPA和FISH在1p缺失的存在与否方面结果一致。在3个OG中,MLPA检测到FISH未显示的19q缺失。对于其他15个OG,MLPA和FISH结果一致。在一个肿瘤占比50%至75%的样本中,MLPA未能检测到FISH显示的1p/19q缺失(尽管具有临界显著性值)。我们得出结论,MLPA是检测福尔马林固定石蜡包埋组织中OG的1p/19q染色体缺失的一种有效且可重复的方法。