Broude N E, Zhang L, Woodward K, Englert D, Cantor C R
Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
Proc Natl Acad Sci U S A. 2001 Jan 2;98(1):206-11. doi: 10.1073/pnas.98.1.206.
We have developed a strategy for multiplex PCR based on PCR suppression. PCR suppression allows DNA target amplification with only one sequence-specific primer per target and a second primer that is common for all targets. Therefore, an n-plex PCR would require only n + 1 primers. We have demonstrated uniform, efficient amplification of targeted sequences in 14-plex PCR. The high specificity of suppression PCR also provides multiplexed amplification with allele specificity. Multiplexed PCR was used to develop assays for genotyping DNA samples from cystic fibrosis-affected individuals. The new approach greatly simplifies primer design, significantly increases the PCR multiplexing level, and decreases the overall primer cost. In addition, this assay is more readily amenable to automation and is therefore suitable for high-throughput genetic diagnostics.
我们基于PCR抑制技术开发了一种多重PCR策略。PCR抑制技术允许每个靶标仅使用一个序列特异性引物和一个所有靶标通用的第二个引物来进行DNA靶标扩增。因此，n重PCR仅需要n + 1个引物。我们已经证明在14重PCR中能够对靶向序列进行均匀、高效的扩增。抑制性PCR的高特异性还能实现具有等位基因特异性的多重扩增。多重PCR被用于开发针对囊性纤维化患者DNA样本进行基因分型的检测方法。这种新方法极大地简化了引物设计，显著提高了PCR多重化水平，并降低了总体引物成本。此外，该检测方法更易于自动化，因此适用于高通量基因诊断。
