Multiplex PCR-based real-time invader assay (mPCR-RETINA): A novel SNP-based method for detecting allellic asymmetries within copy number variation regions
Multiplex PCR-based real-time invader assay (mPCR-RETINA): A novel SNP-based method for detecting allellic asymmetries within copy number variation regions
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DOI：
10.1002/humu.20609
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发表时间：
2008-01-01
期刊：
影响因子：
3.9
通讯作者：
Nakamura, Yusuke
中科院分区：
文献类型：
--
作者：
Hosono, Naoya;Kubo, Michiaki;Nakamura, Yusuke
我们报告了一种结合多重PCR（mPCR-RETINA）的实时入侵检测的发展，这是一种基于SNP的方法，可以测量基因组拷贝数变异（CNV）区域的等位基因比例。RETINA在Invader检测期间监测每个等位基因的实时荧光强度，并检测杂合子个体中由基因组复制/倍增引起的等位基因不对称性。结合mPCR、RETINA和实时定量PCR检测总拷贝数，我们可以估计CNV区域中每个等位基因的拷贝数，这将有助于研究等位基因拷贝数与疾病易感性和药物反应的功能意义。此外，mPCR、RETINA可以有效地细化CNV区域的详细结构。由于RETINA与多重PCR的组合，mPCR-RETINA需要非常少量的基因组DNA用于分析（0.1-0.38 ng/位点）。此外，mPCR RETINA在其简单的方案和靶特异性反应方面具有明显的优势，即使在非独特区域也是如此。我们相信，mPCR，RETINA将提供一个显着的贡献，以确定在CNV区域的功能等位基因。
We report the development of a real-time Invader assay combined with multiplex PCR (mPCR-RETINA), an SNP-based approach that can measure the allelic ratio in copy number variation (CNV) regions of a genome. RETINA monitors the real-time fluorescence intensity of each allele during the Invader assay and detects allelic asymmetries caused by genomic duplication/multiplication in heterozygous individuals. By combining mPCR, RETINA and real-time quantitative PCR that detects total copy number, we can estimate the copy number of each allele in CNV regions, which should be useful for investigating the functional significance of allele copy number with disease susceptibilities and drug responses. Also, mPCR,RETINA can efficiently refine the detailed structures of CNV regions. Due to the combination of RETINA with multiplex PCR, mPCR-RETINA requires a very small amount of genomic DNA for analysis (0.1-0.38 ng/locus). Additionally, mPCR RETINA has clear advantages in its simple protocol and target-specific reaction, even in nonunique regions. We believe mPCR,RETINA will provide a significant contribution to identifying functional alleles in CNV regions.