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서강대학교

Biosens. Bioelectron., 2023 Jun 29;237:115489 | https://doi.org/10.1016/j.bios.2023.115489 Bubble-free diatoms polymerase chain reaction

Yonghee Shin^a,b,c,1, Taejin Kwak ^f,1, Keumrai Whang ^a,1, Yuseung Jo ^a, Jeong Ha Hwang ^a, Inhyeok Hwang ^a, Hyun Ji An ^g, Youngwook Lim ^f, Inhee Choi ^g, Dongchoul Kim ^f, Luke P. Lee ^c,d,e, Taewook Kang ^a,b

^aDepartment of Chemical and Biomolecular Engineering, Sogang University, Seoul, 121-742, South Korea
^bInstitute of Integrated Biotechnology, Sogang University, Seoul, 121-742, South Korea
^cRenal Division and Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA
^dDepartment of Bioengineering, Department of Electrical Engineering and Computer Science, University of California at Berkeley, Berkeley, CA, USA
^eInstitute of Quantum Biophysics, Department of Biophysics, Sungkyunkwan University, Suwon, 16419, South Korea
^fDepartment of Mechanical Engineering, Sogang University, Seoul, 04107, South Korea
^gDepartment of Life Science, University of Seoul, Seoul, 02504, South Korea

¹These authors contributed equally.
Corresponding authors: Dongchoul Kim, Luke P. Lee, Taewook Kang

Abstract

Polymerase chain reaction (PCR) in small fluidic systems not only improves speed and sensitivity of deoxyribonucleic acid (DNA) amplification but also achieves high-throughput quantitative analyses. However, air bubble trapping and growth during PCR has been considered as a critical problem since it causes the failure of DNA amplification. Here we report bubble-free diatom PCR by exploiting a hierarchically porous silica structure of single-celled algae. We show that femtoliters of PCR solution can be spontaneously loaded into the diatom interior without air bubble trapping due to the surface hydrophilicity and pore structure of the diatom. We discover that a large pressure gradient between air bubbles and nanopores rapidly removes residual air bubbles through the periodically arrayed nanopores during thermal cycling. We demonstrate the DNA amplification by diatom PCR without air bubble trapping and growth. Finally, we successfully detect DNA fragments of SARS-CoV-2 with as low as 10 copies/μl by devising a microfluidic device integrated with diatoms assembly. We believe that our work can be applied to many PCR applications for innovative molecular diagnostics and provides new opportunities for naturally abundant diatoms to create innovative biomaterials in real-world applications.

논문정보

형식Research article
게재일2023년 06월 (BRIC 등록일 2023-07-21)
연구진국내(교신)+국외 연구진
분야 바이오·의료융합 > 바이오센싱 및 나노바이오물질

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