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Toward Rapid Detection of Viable Bacteria in Whole Blood for Early Sepsis Diagnostics and Susceptibility Testing
ACS Sensors ( IF 8.9 ) Pub Date : 2021-08-19 , DOI: 10.1021/acssensors.1c01219
Sharath Narayana Iyengar 1, 2 , Jiri Dietvorst 3 , Amparo Ferrer-Vilanova 3 , Gonzalo Guirado 4 , Xavier Muñoz-Berbel 3 , Aman Russom 1, 2
Affiliation  

Sepsis is a serious bloodstream infection where the immunity of the host body is compromised, leading to organ failure and death of the patient. In early sepsis, the concentration of bacteria is very low and the time of diagnosis is very critical since mortality increases exponentially with every hour after infection. Common culture-based methods fail in fast bacteria determination, while recent rapid diagnostic methods are expensive and prone to false positives. In this work, we present a sepsis kit for fast detection of bacteria in whole blood, here achieved by combining selective cell lysis and a sensitive colorimetric approach detecting as low as 103 CFU/mL bacteria in less than 5 h. Homemade selective cell lysis buffer (combination of saponin and sodium cholate) allows fast processing of whole blood in 5 min while maintaining bacteria alive (100% viability). After filtration, retained bacteria on filter paper are incubated under constant illumination with the electrochromic precursors, i.e., ferricyanide and ferric ammonium citrate. Viable bacteria metabolically reduce iron(III) complexes, initiating a photocatalytic cascade toward Prussian blue formation. As a proof of concept, we combine this method with antibiotic susceptibility testing to determine the minimum inhibitory concentration (MIC) using two antibiotics (ampicillin and gentamicin). Although this kit is used to demonstrate its applicability to sepsis, this approach is expected to impact other key sectors such as hygiene evaluation, microbial contaminated food/beverage, or UTI, among others.

中文翻译:

快速检测全血中的活菌,用于早期脓毒症诊断和药敏试验

脓毒症是一种严重的血流感染,宿主机体的免疫力受到损害,导致器官衰竭和患者死亡。在早期败血症中,细菌浓度非常低,诊断时间非常关键,因为感染后每小时死亡率呈指数增长。常见的基于培养的方法无法快速确定细菌,而最近的快速诊断方法昂贵且容易出现假阳性。在这项工作中,我们提出了一种用于快速检测全血中细菌的败血症试剂盒,通过结合选择性细胞裂解和灵敏的比色方法检测低至 10 3CFU/mL 细菌在不到 5 小时内。自制的选择性细胞裂解缓冲液(皂苷和胆酸钠的组合)可在 5 分钟内快速处理全血,同时保持细菌存活(100% 存活率)。过滤后,滤纸上保留的细菌在恒定光照下与电致变色前体,即铁氰化物和柠檬酸铁铵一起温育。活细菌代谢还原铁 (III) 复合物,启动光催化级联反应,形成普鲁士蓝。作为概念证明,我们将此方法与抗生素药敏试验相结合,以确定使用两种抗生素(氨苄青霉素和庆大霉素)的最低抑菌浓度 (MIC)。尽管该试剂盒用于证明其对败血症的适用性,
更新日期:2021-09-24
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