Cultureless enumeration of live bacteria in urinary tract infection by single-cell Raman spectroscopy

Jingkai Wang; Kang Kong; Kang Kong; Chen Guo; Chen Guo; Guangyao Yin; Siyu Meng; Lu Lan; Liqiang Luo; Yizhi Song; Yizhi Song; Yizhi Song

doi:10.3389/fmicb.2023.1144607

Frontiers in Microbiology (Mar 2023)

Cultureless enumeration of live bacteria in urinary tract infection by single-cell Raman spectroscopy

Jingkai Wang,
Kang Kong,
Kang Kong,
Chen Guo,
Chen Guo,
Guangyao Yin,
Siyu Meng,
Lu Lan,
Liqiang Luo,
Yizhi Song,
Yizhi Song,
Yizhi Song

Affiliations

Jingkai Wang: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
Kang Kong: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
Kang Kong: Department of Chemistry, College of Sciences, Shanghai University, Shanghai, China
Chen Guo: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
Chen Guo: Division of Life Sciences and Medicine, School of Biomedical Engineering (Suzhou), University of Science and Technology of China, Suzhou, China
Guangyao Yin: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
Siyu Meng: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
Lu Lan: VibroniX, Inc., Suzhou, China
Liqiang Luo: Department of Chemistry, College of Sciences, Shanghai University, Shanghai, China
Yizhi Song: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
Yizhi Song: Division of Life Sciences and Medicine, School of Biomedical Engineering (Suzhou), University of Science and Technology of China, Suzhou, China
Yizhi Song: Chongqing Guoke Medical Technology Development Co., Ltd., Chongqing, China

DOI: https://doi.org/10.3389/fmicb.2023.1144607
Journal volume & issue: Vol. 14

Abstract

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Urinary tract infections (UTIs) are the most common outpatient infections. Obtaining the concentration of live pathogens in the sample is crucial for the treatment. Still, the enumeration depends on urine culture and plate counting, which requires days of turn-around time (TAT). Single-cell Raman spectra combined with deuterium isotope probing (Raman-DIP) has been proven to identify the metabolic-active bacteria with high accuracy but is not able to reveal the number of live pathogens due to bacteria replication during the Raman-DIP process. In this study, we established a new approach of using sodium acetate to inhibit the replication of the pathogen and applying Raman-DIP to identify the active single cells. By combining microscopic image stitching and recognition, we could further improve the efficiency of the new method. Validation of the new method on nine artificial urine samples indicated that the exact number of live pathogens obtained with Raman-DIP is consistent with plate-counting while shortening the TAT from 18 h to within 3 h, and the potential of applying Raman-DIP for pathogen enumeration in clinics is promising.

Published in Frontiers in Microbiology

ISSN: 1664-302X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: http://www.frontiersin.org/journals/microbiology

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