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采用克隆竞争物的简单通用的分子拷贝数测量方法。

Simple and versatile molecular method of copy-number measurement using cloned competitors.

机构信息

Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Korea.

出版信息

PLoS One. 2013 Jul 30;8(7):e69414. doi: 10.1371/journal.pone.0069414. Print 2013.

DOI:10.1371/journal.pone.0069414
PMID:23936009
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3728337/
Abstract

Variations and alterations of copy numbers (CNVs and CNAs) carry disease susceptibility and drug responsiveness implications. Although there are many molecular methods to measure copy numbers, sensitivity, reproducibility, cost, and time issues remain. In the present study, we were able to solve those problems utilizing our modified real competitive PCR method with cloned competitors (mrcPCR). First, the mrcPCR for ERBB2 copy number was established, and the results were comparable to current standard methods but with a shorter assay time and a lower cost. Second, the mrcPCR assays for 24 drug-target genes were established, and the results in a panel of NCI-60 cells were comparable to those from real-time PCR and microarray. Third, the mrcPCR results for FCGR3A and the FCGR3B CNVs were comparable to those by the paralog ratio test (PRT), but without PRT's limitations. These results suggest that mrcPCR is comparable to the currently available standard or the most sensitive methods. In addition, mrcPCR would be invaluable for measurement of CNVs in genes with variants of similar structures, because combination of the other methods is not necessary, along with its other advantages such as short assay time, small sample amount requirement, and applicability to all sequences and genes.

摘要

拷贝数变异（CNVs 和 CNAs）的变化和改变与疾病易感性和药物反应性有关。虽然有许多分子方法可以测量拷贝数，但仍然存在灵敏度、重现性、成本和时间问题。在本研究中，我们能够利用我们改良的带有克隆竞争物的实时竞争 PCR 方法（mrcPCR）解决这些问题。首先，建立了 ERBB2 拷贝数的 mrcPCR，结果与当前标准方法相当，但检测时间更短，成本更低。其次，建立了 24 个药物靶基因的 mrcPCR 检测方法，在 NCI-60 细胞系中的结果与实时 PCR 和微阵列相当。第三，FCGR3A 和 FCGR3B CNVs 的 mrcPCR 结果与等位基因比测试（PRT）相当，但没有 PRT 的局限性。这些结果表明，mrcPCR 与当前可用的标准或最敏感的方法相当。此外，mrcPCR 对于测量具有相似结构变体的基因中的 CNVs 非常有价值，因为不需要结合其他方法，并且具有检测时间短、样品量需求小以及适用于所有序列和基因等优点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/c7f2610815d0/pone.0069414.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/66124774aa41/pone.0069414.g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/742b5d03e0b2/pone.0069414.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/228c0e7f6574/pone.0069414.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/2569a21aaa44/pone.0069414.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/c3b353df16a3/pone.0069414.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/730ca4e7856b/pone.0069414.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/c7f2610815d0/pone.0069414.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/66124774aa41/pone.0069414.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/6c1a230ca34b/pone.0069414.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/742b5d03e0b2/pone.0069414.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/228c0e7f6574/pone.0069414.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/2569a21aaa44/pone.0069414.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/c3b353df16a3/pone.0069414.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/730ca4e7856b/pone.0069414.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1ea/3728337/c7f2610815d0/pone.0069414.g008.jpg

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