Onsite detection of airborne antibiotic-resistant bacteria via Cas9 nickase-triggered amplification reactions

IF 11.3 1区环境科学与生态学 Q1 ENGINEERING, ENVIRONMENTAL

Journal of Hazardous Materials Pub Date : 2025-09-05 Epub Date: 2025-06-06 DOI:10.1016/j.jhazmat.2025.138850

Seung Beom Seo , Jina Lee , Sangsoo Choi , Dongmin Shin , Soojin Jang , Yeonwoo Jeong , Seong Uk Son , Taejoon Kang , Juyeon Jung , Kyujung Kim , Jungho Hwang , Eun-Kyung Lim

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引用次数: 0

Abstract

Antibiotic resistance is a critical global health issue, with methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) being major pathogens causing pneumonia and sepsis. In this study, we introduce the Cas9 nickase-triggered amplification reaction (CN-TAR) assay - onsite, real-time detection method designed to help prevent airborne transmission of these pathogens. The assay utilizes Cas9 nickase to specifically cleave target DNA, followed by rolling circle amplification for single-step detection. To enhance filed applicability, a portable isothermal PCR device was integrated into the system. The CN-TAR assay was validated using synthetic nucleic acids, cultured bacteria, and airborne samples, achieving detection limits of 1.40 copies/μL for MRSA and 1.13 copies/μL for VRE. It demonstrated high sensitivity and rapid turnaround time. Furthermore, its performance was comparable to that of conventional reverse transcription PCR (RT-PCR), confirming its reliability for airborne antibiotic-resistant bacteria monitoring. This study presents a practical on-site detection platform, and the results highlight the CN-TAR assay as a promising tool for real-time surveillance and detection, contributing to effective infection control and public health safety.

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Cas9镍酶触发扩增反应现场检测空气中耐药细菌

抗生素耐药性是一个严重的全球卫生问题，耐甲氧西林金黄色葡萄球菌（MRSA）和耐万古霉素肠球菌（VRE）是导致肺炎和败血症的主要病原体。在本研究中，我们引入了Cas9镍酶触发扩增反应（CN-TAR）检测-现场实时检测方法，旨在帮助预防这些病原体的空气传播。该方法利用Cas9缺口酶特异性切割靶DNA，然后进行滚圈扩增，进行单步检测。为了提高现场适用性，在系统中集成了便携式等温PCR装置。通过合成核酸、培养细菌和空气样品对CN-TAR法进行验证，MRSA和VRE的检出限分别为1.40 copies/μL和1.13 copies/μL。该方法灵敏度高，周转时间短。此外，其性能可与传统的逆转录PCR （RT-PCR）相媲美，证实了其在空气中耐药菌监测中的可靠性。本研究提出了一个实用的现场检测平台，结果表明，CN-TAR检测是一种有前途的实时监测和检测工具，有助于有效控制感染和公共卫生安全。

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来源期刊

Journal of Hazardous Materials 工程技术-工程：环境

CiteScore

25.40

自引率

5.90%

发文量

3059

审稿时长

58 days

期刊介绍： The Journal of Hazardous Materials serves as a global platform for promoting cutting-edge research in the field of Environmental Science and Engineering. Our publication features a wide range of articles, including full-length research papers, review articles, and perspectives, with the aim of enhancing our understanding of the dangers and risks associated with various materials concerning public health and the environment. It is important to note that the term "environmental contaminants" refers specifically to substances that pose hazardous effects through contamination, while excluding those that do not have such impacts on the environment or human health. Moreover, we emphasize the distinction between wastes and hazardous materials in order to provide further clarity on the scope of the journal. We have a keen interest in exploring specific compounds and microbial agents that have adverse effects on the environment.