김한솔 (KAIST, KRIBB) | 한빛사논문 > 한빛사

한빛사논문

조회374

김한솔Hansol Kim

KAIST, KRIBB

CV File 209 kb

Biosens. Bioelectron., Feb 20 2024, 253 S0956-5663(24)00152-0 | https://doi.org/10.1016/j.bios.2024.116147 Multifunctional self-priming hairpin probe-based isothermal nucleic acid amplification and its applications for COVID-19 diagnosis

Hansol Kim ^a,b, Seoyoung Lee ^a, Yong Ju ^a, Hyoyong Kim ^a, Hyowon Jang ^a,b, Yeonkyung Park ^a,b, Sang Mo Lee ^a, Dongeun Yong ^c, Taejoon Kang ^b,d, Hyun Gyu Park ^a

^aDepartment of Chemical and Biomolecular Engineering (BK 21 four), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea

^bBionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea

^cDepartment of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea

^dSchool of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, Gyeonggi-do, 16419, Republic of Korea

Corresponding authors: Taejoon Kang, Hyun Gyu Park

Abstract

We herein present a multifunctional self-priming hairpin probe-based isothermal amplification, termed MSH, enabling one-pot detection of target nucleic acids. The sophisticatedly designed multifunctional self-priming hairpin (MSH) probe recognizes the target and rearranges to prime itself, triggering the amplification reaction powered by the continuously repeated extension, nicking, and target recycling. As a consequence, a large number of double-stranded DNA (dsDNA) amplicons are produced that could be monitored in real-time using a dsDNA-intercalating dye. Based on this unique design approach, the nucleocapsid (N) and the open reading frame 1 ab (ORF1ab) genes of SARS-CoV-2 were successfully detected down to 1.664 fM and 0.770 fM, respectively. The practical applicability of our method was validated by accurately diagnosing 60 clinical samples with 93.33% sensitivity and 96.67% specificity. This isothermal one-pot MSH technique holds great promise as a point-of-care testing protocol for the reliable detection of a wide spectrum of pathogens, particularly in resource-limited settings.

논문정보

형식Research article
게재일2024년 02월 (BRIC 등록일 2024-03-11)
연구진국내 연구진
분야 생명과학 > 생물공학

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