Enhancing the efficiency of a PCR using gold nanoparticles.
Enhancing the efficiency of a PCR using gold nanoparticles.
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DOI：
10.1093/nar/gni183
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发表时间：
2005-11-27
影响因子：
14.9
通讯作者：
Liu HS
中科院分区：
文献类型：
--
作者：
Li M;Lin YC;Wu CC;Liu HS
我们发现Au纳米颗粒可以显着增强PCR。在PCR试剂中加入0.7 nM的13 nm Au纳米颗粒，PCR效率提高。特别是当保持相同或更高的扩增产率时，可以缩短反应时间，并且可以增加加热/冷却速率。纳米颗粒的优良传热性能应该是提高PCR效率的主要因素。本研究比较了不同的PCR体系、DNA聚合酶、DNA大小和复杂样品。我们的研究结果表明，Au纳米颗粒在较慢的PCR系统（即常规PCR）中将PCR检测的灵敏度提高了5至10倍，在较快的PCR系统（即实时PCR）中提高了至少104倍。在PCR时间缩短一半后，在加入胶体金的实时PCR中可检测到100拷贝/μl的DNA，然而，使用不含胶体金的PCR试剂至少需要106拷贝/μl的DNA才能达到可检测的信号水平。该创新可以使用不昂贵的聚合酶和通用PCR试剂来提高PCR效率。利用纳米粒子提高PCR效率，缩短反应时间，是PCR研究的一个新观点。
We found that the PCR could be dramatically enhanced by Au nanoparticles. With the addition of 0.7 nM of 13 nm Au nanoparticles into the PCR reagent, the PCR efficiency was increased. Especially when maintaining the same or higher amplification yields, the reaction time could be shortened, and the heating/cooling rates could be increased. The excellent heat transfer property of the nanoparticles should be the major factor in improving the PCR efficiency. Different PCR systems, DNA polymerases, DNA sizes and complex samples were compared in this study. Our results demonstrated that Au nanoparticles increase the sensitivity of PCR detection 5- to 10-fold in a slower PCR system (i.e. conventional PCR) and at least 104-fold in a quicker PCR system (i.e. real-time PCR). After the PCR time was shortened by half, the 100 copies/µl DNA were detectable in real-time PCR with gold colloid added, however, at least 106 copies/µl of DNA were needed to reach a detectable signal level using the PCR reagent without gold colloid. This innovation could improve the PCR efficiency using non-expensive polymerases, and general PCR reagent. It is a new viewpoint in PCR, that nanoparticles can be used to enhance PCR efficiency and shorten reaction times.
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