Automatic analysis of multiplex ligation-dependent probe amplification products (exemplified by a commercial kit for prenatal aneuploidy detection)
Automatic analysis of multiplex ligation-dependent probe amplification products (exemplified by a commercial kit for prenatal aneuploidy detection)
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DOI：
10.1002/elps.200500390
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发表时间：
2005-11-01
期刊：
影响因子：
2.9
通讯作者：
Bryndorf, T
中科院分区：
文献类型：
--
作者：
Gerdes, T;Kirchhoff, M;Bryndorf, T
为了在常规产前诊断中使用，我们开发了基于商业多重连接依赖性探针扩增（MLPA）试剂盒的自动非整倍体检测软件和方法。软件和方法确保了可靠、客观和快速的工作流程，并且可以应用于其他类型的MLPA试剂盒。MLPA扩增产物的CE后，软件自动识别每个探针的峰面积，相对于供试品的相邻峰面积进行归一化，计算相对于由正常样品创建的参考的比值，并补偿归一化程序的副作用的比值，该归一化程序根据存在的非整倍性的种类将所有染色体正常DNA峰面积略微放大或缩小。对于染色体13、18、21、X和Y，计算探针可靠性加权平均比值和相应的SD，并检验比值1.0左右超出参考区间的显著性。p
For use in routine prenatal diagnostics, we developed software and methods for automatic aneuploidy detection based on a commercial multiplex ligation-dependent probe amplification (MLPA) kit. Software and methods ensure a reliable, objective, and fast workflow, and may be applied to other types of MLPA kits. Following CE of MLPA amplification products, the software automatically identified the peak area for each probe, normalized it in relation to the neighboring peak areas of the test sample, computed the ratio relative to a reference created from normal samples, and compensated the ratio for a side effect of the normalization procedure that scaled all chromosomally normal DNA peak areas slightly up or down depending on the kind of aneuploidy present. For the chromosomes 13, 18, 21, X, and Y, probe reliability weighted mean ratio values and corresponding SDs were calculated, and the significance for being outside a reference interval around ratio 1.0 was tested. p