Micro-flow cell washing technique combined with single-cell Raman spectroscopy for rapid and automatic antimicrobial susceptibility test of pathogen in urine
Copyright © 2024 Elsevier B.V. All rights reserved..
Facing the rapid spread of antimicrobial resistance, methods based on single-cell Raman spectroscopy have proven their advances in reducing the turn-around time (TAT) of antimicrobial susceptibility tests (AST). However, the Raman-based methods are still hindered by the prolonged centrifugal cell washing procedure, which may require complex labor operation and induce high mechanical stress, resulting in a pretreatment time of over 1 h as well as a high cell-loss probability. In this study, we developed a micro-flow cell washing device and corresponding Raman-compatible washing chips, which were able to automatically remove the impurities in the samples, retain the bacterial cell and perform Raman spectra acquisition in situ. Results of washing the 5- and 10-μm polymethyl methacrylate (PMMA) microspheres showed that the novel technique achieved a successful removal of 99 % impurity and an 80 % particle retention rate after 6 to 10 cycles of washing. The micro-flow cell washing technique could complete the pretreatment for urine samples in a 96-well plate within 10 min, only taking 15 % of the handling time required by centrifugation. The AST profiles of urine sample spiked with E. coli 25922, E. faecalis 29212, and S. aureus 29213 obtained by the proposed Raman-based approach were found to be 100 % consistent with the results from broth micro-dilution while reducing the TAT to 3 h from several days which is required by the latter. Our study has demonstrated the micro-flow cell washing technique is a reliable, fast and compatible approach to replace centrifuge washing for sample pretreatment of Raman-AST and could be readily applied in clinical scenarios.
| Media Type: |
Electronic Article |
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| Year of Publication: |
2024 |
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| Publication: |
2024 |
| Contained In: |
To Main Record - volume:277 |
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| Contained In: |
Talanta - 277(2024) vom: 01. Juli, Seite 126354 |
| Language: |
English |
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| Contributors: |
Zhao, Luoqi [Author] |
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| Links: |
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| Keywords: |
9011-14-7 |
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| Notes: |
Date Completed 18.07.2024 Date Revised 18.07.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.talanta.2024.126354 |
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| funding: |
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| Supporting institution / Project title: |
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NLM373389736 |
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| 520 | |a Facing the rapid spread of antimicrobial resistance, methods based on single-cell Raman spectroscopy have proven their advances in reducing the turn-around time (TAT) of antimicrobial susceptibility tests (AST). However, the Raman-based methods are still hindered by the prolonged centrifugal cell washing procedure, which may require complex labor operation and induce high mechanical stress, resulting in a pretreatment time of over 1 h as well as a high cell-loss probability. In this study, we developed a micro-flow cell washing device and corresponding Raman-compatible washing chips, which were able to automatically remove the impurities in the samples, retain the bacterial cell and perform Raman spectra acquisition in situ. Results of washing the 5- and 10-μm polymethyl methacrylate (PMMA) microspheres showed that the novel technique achieved a successful removal of 99 % impurity and an 80 % particle retention rate after 6 to 10 cycles of washing. The micro-flow cell washing technique could complete the pretreatment for urine samples in a 96-well plate within 10 min, only taking 15 % of the handling time required by centrifugation. The AST profiles of urine sample spiked with E. coli 25922, E. faecalis 29212, and S. aureus 29213 obtained by the proposed Raman-based approach were found to be 100 % consistent with the results from broth micro-dilution while reducing the TAT to 3 h from several days which is required by the latter. Our study has demonstrated the micro-flow cell washing technique is a reliable, fast and compatible approach to replace centrifuge washing for sample pretreatment of Raman-AST and could be readily applied in clinical scenarios | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Antimicrobial susceptibility test | |
| 650 | 4 | |a Cell washing | |
| 650 | 4 | |a Micro-flow | |
| 650 | 4 | |a Single-cell Raman spectroscopy | |
| 650 | 7 | |a Anti-Bacterial Agents |2 NLM | |
| 650 | 7 | |a Polymethyl Methacrylate |2 NLM | |
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| 700 | 1 | |a Jiang, Zheng |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Jingkai |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Xinyue |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Zhiqiang |e verfasserin |4 aut | |
| 700 | 1 | |a Hu, Huijie |e verfasserin |4 aut | |
| 700 | 1 | |a Qi, Xiangdong |e verfasserin |4 aut | |
| 700 | 1 | |a Zeng, Huan |e verfasserin |4 aut | |
| 700 | 1 | |a Song, Yizhi |e verfasserin |4 aut | |
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