Goo Nam-In, Kim Dong-Eun
Department of Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029 Korea.
Biochip J. 2016;10(4):262-271. doi: 10.1007/s13206-016-0402-6. Epub 2016 Jul 29.
Rolling circle amplification (RCA) developed in the mid-1990s has been widely used as an efficient isothermal DNA amplification process for molecular diagnosis. This enzymatic process amplifies target DNA sequences with high fidelity and specificity by using the strand displacing DNA polymerases. The product of RCA is long single-stranded DNA that contains tandem repeat of target sequence. Isothermal reaction amplification condition of RCA has an advantage over conventional polymerase chain reaction, because no temperature cycling devices are needed for RCA. Thus, RCA is suitable tool for point-of-care detection of target nucleic acids as well as facile detection of target genes. Combined with various detection methods, RCA could amplify and detect femtomolar scale of target nucleic acids with a specificity of one or two base discrimination. Herein, RCA technology is reviewed with an emphasis on molecular diagnosis of microRNAs, infectious pathogens, and point mutations.
20世纪90年代中期开发的滚环扩增（RCA）已被广泛用作分子诊断中一种高效的等温DNA扩增方法。该酶促过程通过使用链置换DNA聚合酶以高保真度和特异性扩增靶DNA序列。RCA的产物是包含靶序列串联重复的长单链DNA。RCA的等温反应扩增条件优于传统的聚合酶链反应，因为RCA不需要温度循环装置。因此，RCA是用于即时检测靶核酸以及轻松检测靶基因的合适工具。与各种检测方法相结合，RCA可以扩增和检测飞摩尔级的靶核酸，特异性可达一两个碱基区分。本文对RCA技术进行了综述，重点介绍了其在微小RNA、传染性病原体和点突变分子诊断中的应用。
