Pneumatic Nano-Sieve for CRISPR-based Detection of Drug-resistant Bacteria.
Pneumatic Nano-Sieve for CRISPR-based Detection of Drug-resistant Bacteria.
复制标题
用于基于 CRISPR 检测耐药细菌的气动纳米筛。
DOI：
10.1101/2023.08.17.553737
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Du,Ke
中科院分区：
文献类型：
--
作者：
Peng,Ruonan;Chen,Xinye;Xu,Fengjun;Hailstone,Richard;Men,Yujie;Du,Ke
耐甲氧西林金黄色葡萄球菌（MRSA）等耐药性细菌感染的日益普遍，已成为一个重大的公共卫生问题。及时检测MRSA对于及时进行医疗干预、限制其传播和减少抗生素耐药性至关重要。在这里，我们介绍了一种小型化的纳米筛装置，其特征在于具有可调节的腔室，用于从人血浆样品中高效纯化MRSA。通过使用填充的磁珠作为过滤器并利用纳米筛通道的可变形性，我们实现了MRSA的15倍的芯片上浓缩因子。我们将该设备与重组酶聚合酶扩增（RPA）和成簇的规则间隔短回文重复序列（CRISPR）-Cas检测系统集成在一起，导致芯片上检测限（LOD）约为100 CFU mL−1。这种开发的方法提供了一种快速，精确和无离心机的解决方案，适用于床旁诊断，有可能显着改善资源有限的医疗条件下的患者预后。
The increasing prevalence of antibiotic-resistant bacterial infections, particularly methicillin-resistant Staphylococcus aureus (MRSA), presents a significant public health concern. Timely detection of MRSA is crucial to enable prompt medical intervention, limit its spread, and reduce antimicrobial resistance. Here, we introduce a miniaturized nano-sieve device featuring a pneumatically-regulated chamber for highly efficient MRSA purification from human plasma samples. By using packed magnetic beads as a filter and leveraging the deformability of the nano-sieve channel, we achieved an on-chip concentration factor of ∼15-fold for MRSA. We integrated this device with recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas detection system, resulting in an on-chip limit of detection (LOD) of approximately 100 CFU mL−1. This developed approach provides a rapid, precise, and centrifuge-free solution suitable for point-of-care diagnostics, with the potential to significantly improve patient outcomes in resource-limited medical conditions.