A Portable Droplet Magnetofluidic Device for Point-of-Care Detection of Multidrug-Resistant Candida auris.

A Portable Droplet Magnetofluidic Device for Point-of-Care Detection of Multidrug-Resistant Candida auris.
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一种可移植的液滴磁磁体装置,用于耐多药念珠菌的护理点检测。

DOI:
10.3389/fbioe.2022.826694
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发表时间:
2022
影响因子:
5.7
通讯作者:
Wang, Tza-Huei
Wang, Tza-Huei
中科院分区:
工程技术2区
文献类型:
--
作者:
Lee, Pei-Wei;Totten, Marissa;Chen, Liben;Chen, Fan-En;Trick, Alexander Y.;Shah, Kushagra;Ngo, Hoan Thanh;Jin, Mei;Hsieh, Kuangwen;Zhang, Sean X.;Wang, Tza-Huei

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耳念珠菌是一种新兴的多重耐药真菌病原体,可引起严重和致命的感染。迄今为止,C.在五大洲三十三个国家的医疗机构中,auris引发了疫情。为了控制和潜在地防止其传播,迫切需要能够快速筛查患者、密切患者接触者和监测环境来源的即时(POC)诊断。液滴磁流体(DM),它利用核酸结合磁珠实现POC-顺从的核酸检测平台,提供了一个有前途的解决方案。在此,我们报告了第一个用于C.耳。作为POC.auris的一部分,我们已经整合了一个手持式细胞裂解模块,可以裂解C。耳细胞2分钟的动手时间。随后,在手掌大小的自动DM设备中,C。耳和对照DNA通过电动磁力臂进行磁力提取和纯化,最后通过小型化快速PCR模块和小型化荧光检测器通过双重实时定量PCR测定进行扩增-所有操作均在≤30 min内完成。为了演示,我们使用POC.auris检测C。从3个主要分支的耳念珠菌分离株,与其他念珠菌属没有交叉反应性,检测限为1000个菌落形成单位/mL。总之,POC.auris提供了一个潜在的有用的工具,用于打击C。耳。
Candida auris is an emerging multidrug-resistant fungal pathogen that can cause severe and deadly infections. To date, C. auris has spurred outbreaks in healthcare settings in thirty-three countries across five continents. To control and potentially prevent its spread, there is an urgent need for point-of-care (POC) diagnostics that can rapidly screen patients, close patient contacts, and surveil environmental sources. Droplet magnetofluidics (DM), which leverages nucleic acid-binding magnetic beads for realizing POC-amenable nucleic acid detection platforms, offers a promising solution. Herein, we report the first DM device—coined POC.auris—for POC detection of C. auris. As part of POC.auris, we have incorporated a handheld cell lysis module that lyses C. auris cells with 2 min hands-on time. Subsequently, within the palm-sized and automated DM device, C. auris and control DNA are magnetically extracted and purified by a motorized magnetic arm and finally amplified via a duplex real-time quantitative PCR assay by a miniaturized rapid PCR module and a miniaturized fluorescence detector—all in ≤30 min. For demonstration, we use POC.auris to detect C. auris isolates from 3 major clades, with no cross reactivity against other Candida species and a limit of detection of ∼300 colony forming units per mL. Taken together, POC.auris presents a potentially useful tool for combating C. auris.
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