Kim Jinjoo, Kim Hansol, Park Ji Ho, Jon Sangyong
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Daejeon 34141, Republic of Korea.
KAIST Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Daejeon 34141, Republic of Korea.
Nanotheranostics. 2017 May 6;1(2):178-185. doi: 10.7150/ntno.18720. eCollection 2017.
The polymerase chain reaction (PCR) has been an essential tool for diagnosis of infectious diseases, but conventional PCR still has some limitations with respect to applications to point-of-care (POC) diagnostic systems that require rapid detection and miniaturization. Here we report a light-based PCR method, termed as photo-PCR, which enables rapid detection of bacteria in a single step. In the photo-PCR system, poly(enthylene glycol)-modified gold nanorods (PEG-GNRs), used as a heat generator, are added into the PCR mixture, which is subsequently periodically irradiated with a 808-nm laser to create thermal cycling. Photo-PCR was able to significantly reduce overall thermal cycling time by integrating bacterial cell lysis and DNA amplification into a single step. Furthermore, when combined with KAPA2G fast polymerase and cooling system, the entire process of bacterial genomic DNA extraction and amplification was further shortened, highlighting the potential of photo-PCR for use in a portable, POC diagnostic system.
聚合酶链反应(PCR)一直是诊断传染病的重要工具,但传统PCR在应用于需要快速检测和小型化的即时检测(POC)诊断系统方面仍存在一些局限性。在此,我们报告一种基于光的PCR方法,称为光PCR,它能够一步快速检测细菌。在光PCR系统中,用作发热体的聚乙二醇修饰的金纳米棒(PEG-GNRs)被添加到PCR混合物中,随后用808纳米激光对其进行周期性照射以产生热循环。光PCR能够通过将细菌细胞裂解和DNA扩增整合为一步,显著缩短整个热循环时间。此外,当与KAPA2G快速聚合酶和冷却系统结合使用时,细菌基因组DNA提取和扩增的整个过程进一步缩短,突出了光PCR在便携式POC诊断系统中的应用潜力。