Rapid ultra-sensitive diagnosis of clostridium difficile infection using a SERS-based lateral flow assay
Rapid ultra-sensitive diagnosis of clostridium difficile infection using a SERS-based lateral flow assay
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DOI：
10.1039/d1an00726b
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发表时间：
2021-06-21
期刊：
影响因子：
4.2
通讯作者：
Graham, Duncan
中科院分区：
文献类型：
--
作者：
Hassanain, Waleed A.;Spoors, Julia;Graham, Duncan
艰难梭菌（C. diff）感染是与住院患者的高发病率和死亡率相关的最具传染性的疾病之一。准确的诊断可以通过确定最有效的治疗方法来减缓其传播。在此，我们报告了一种新的测试平台，作为选择性，灵敏，快速和成本效益的诊断C的概念验证。基于双重测量的差异感染（CDI）。这是通过使用基于表面增强拉曼散射的侧向流测定（基于SERS的LFA）检测两种特异性生物标志物，表面层蛋白A（SlpA）和毒素B（ToxB）来实现的。以前还没有描述SlpA与ToxB的同时双重检测用于CDI的临床诊断。SlpA生物标志物“AKDGSTKEDQLVDALA”于2018年首次由我们的团队作为物种特异性鉴定工具报道。第二个生物标志物ToxB是C.不同的致病菌株，并需要确认真正的感染致病性。所提出的基于SERS的LFA平台能够在20分钟内使用双链LF测试条在单独的测试线上快速双链检测SlpA和ToxB。使用手持式拉曼光谱仪扫描测试线允许两种生物标志物的高灵敏度定量检测，最低可观察浓度为0.01 pg μ L-1。在这种基于SERS的LFA中使用手持设备而不是台式机器，为在护理点（POC）快速，选择性，灵敏和廉价的CDI临床评价铺平了道路，样品积压最少。
Clostridium difficile (C. diff) infection is one of the most contagious diseases associated with high morbidity and mortality rates in hospitalised patients. Accurate diagnosis can slow its spread by determining the most effective treatment. Herein, we report a novel testing platform as a proof-of-concept for the selective, sensitive, rapid and cost-effective diagnosis of C. diff infection (CDI) based on a duplex measurement. This was achieved by detecting two specific biomarkers, surface layer protein A (SlpA) and toxin B (ToxB), using a surface enhanced Raman scattering-based lateral flow assay (SERS-based LFA). The simultaneous duplex detection of SlpA with ToxB has not been described for the clinical diagnosis of CDI previously. The SlpA biomarker "AKDGSTKEDQLVDALA" was first reported by our group in 2018 as a species-specific identification tool. The second biomarker, ToxB, is the essential virulence biomarker of C. diff pathogenic strains and is required to confirm true infection pathogenicity. The proposed SERS-based LFA platform enabled rapid duplex detection of SlpA and ToxB on separate test lines using a duplex LF test strip within 20 minutes. The use of a handheld Raman spectrometer to scan test lines allowed for the highly sensitive quantitative detection of both biomarkers with a lowest observable concentration of 0.01 pg mu L-1. The use of a handheld device in this SERS-based LFA instead of benchtop machine paves the way for rapid, selective, sensitive and cheap clinical evaluation of CDI at the point of care (POC) with minimal sample backlog.