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울산과학기술원 (UNIST)

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Biosens. Bioelectron., Nov 22 2024, 271 S0956-5663(24)00957-6 | https://doi.org/10.1016/j.bios.2024.116950 Fast and accurate multi-bacterial identification using cleavable and FRET-based peptide nucleic acid probes

Sungho Kim ^a,1, Hwi Hyun ^a,1, Jae-Kyeong Im ^a, Min Seok Lee ^a, Hwasoo Koh ^a, Donghoon Kang ^a, Si-Hyeong Nho ^a, Joo H. Kang ^a, Taejoon Kwon ^a,b, Hajin Kim ^a,b

^aDepartment of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 44919, Republic of Korea
^bCenter for Genomic Integrity, Institute for Basic Science, Ulsan, 44919, Republic of Korea

¹These authors contributed equally to this work.
Corresponding authors: Joo H. Kang, Taejoon Kwon, Hajin Kim

Abstract

Fast and accurate identification of pathogenic microbes in patient samples is crucial for the timely treatment of acute infectious diseases such as sepsis. The fluorescence in situ hybridization (FISH) technique allows the rapid detection and identification of microbes based on their variation in genomic sequence without time-consuming culturing or sequencing. However, the recent explosion of microbial genomic data has made it challenging to design an appropriate set of probes for microbial mixtures. We developed a novel set of peptide nucleic acid (PNA)-based FISH probes with optimal target specificity by analyzing the variations in 16S ribosomal RNA sequence across all bacterial species. Owing to their superior penetration into bacteria and higher mismatch sensitivity, the PNA probes distinguished seven bacterial species commonly observed in bacteremia with 96-99.9% accuracy using our optimized FISH procedure. Detection based on Förster resonance energy transfer (FRET) between pairs of adjacent binding PNA probes eliminated crosstalk between species. Rapid sequential species identification was implemented, using chemically cleavable fluorophores, without compromising detection accuracy. Owing to their outstanding accuracy and enhanced speed, this set of techniques shows great potential for clinical use.

논문정보

형식Research article
게재일2024년 11월 (BRIC 등록일 2024-12-09)
연구진국내 연구진
분야 생명과학 > 분자생물학

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