Song Younseong, Park Nahyun, Jo Da Ae, Kim Jueun, Yong Dongeun, Song Jayeon, Park Yoo Min, Lee Seok Jae, Kim Yong Tae, Im Sung Gap, Choi Bong Gill, Kang Taejoon, Lee Kyoung G
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 34141, Republic of Korea.
Center for Nano Bio Development, National Nanofab Center (NNFC), Daejeon, 34141, Republic of Korea.
Nano Converg. 2023 May 27;10(1):25. doi: 10.1186/s40580-023-00370-w.
Sensitive and accurate capture, enrichment, and identification of drug-resistant bacteria on human skin are important for early-stage diagnosis and treatment of patients. Herein, we constructed a three-dimensional hierarchically structured polyaniline nanoweb (3D HPN) to capture, enrich, and detect drug-resistant bacteria on-site by rubbing infected skins. These unique hierarchical nanostructures enhance bacteria capture efficiency and help severely deform the surface of the bacteria entrapped on them. Therefore, 3D HPN significantly contributes to the effective and reliable recovery of drug-resistant bacteria from the infected skin and the prevention of potential secondary infection. The recovered bacteria were successfully identified by subsequent real-time polymerase chain reaction (PCR) analysis after the lysis process. The molecular analysis results based on a real-time PCR exhibit excellent sensitivity to detecting target bacteria of concentrations ranging from 10 to 10 CFU/mL without any fluorescent signal interruption. To confirm the field applicability of 3D HPN, it was tested with a drug-resistant model consisting of micropig skin similar to human skin and Klebsiella pneumoniae carbapenemase-producing carbapenem-resistant Enterobacteriaceae (KPC-CRE). The results show that the detection sensitivity of this assay is 10 CFU/mL. Therefore, 3D HPN can be extended to on-site pathogen detection systems, along with rapid molecular diagnostics through a simple method, to recover KPC-CRE from the skin.
对人皮肤上耐药菌进行灵敏、准确的捕获、富集和鉴定，对于患者的早期诊断和治疗至关重要。在此，我们构建了一种三维分层结构的聚苯胺纳米网（3D HPN），通过擦拭感染皮肤来现场捕获、富集和检测耐药菌。这些独特的分层纳米结构提高了细菌捕获效率，并使捕获在其上的细菌表面严重变形。因此，3D HPN对从感染皮肤中有效、可靠地回收耐药菌以及预防潜在的二次感染有显著贡献。在裂解过程后，通过后续的实时聚合酶链反应（PCR）分析成功鉴定了回收的细菌。基于实时PCR的分子分析结果对浓度范围为10至10 CFU/mL的目标细菌检测具有出色的灵敏度，且无任何荧光信号中断。为了确认3D HPN的现场适用性，我们使用了由类似于人类皮肤的微型猪皮肤和产肺炎克雷伯菌碳青霉烯酶的耐碳青霉烯肠杆菌科细菌（KPC-CRE）组成的耐药模型进行测试。结果表明，该检测方法的检测灵敏度为10 CFU/mL。因此，3D HPN可以扩展到现场病原体检测系统，通过一种简单的方法进行快速分子诊断，从皮肤中回收KPC-CRE。
