Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol
Usually, 18–48 h are needed for the identification of microbial pathogens causing urinary tract infections (UTIs) by urine culture. Moreover, antimicrobial susceptibility testing (AST) takes an additional 18–24 h. Rapid identification and AST of the pathogens allow fast and precise treatment. The ob...
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doaj-596213cff26d4c5cb4eddaa716e3cc602020-11-25T00:39:35ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-06-011010.3389/fmicb.2019.01182458039Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined ProtocolWei Li0Enhua Sun1Ying Wang2Hongwei Pan3Yi Zhang4Yong Li5Xin Zhang6Chen Li7Lutao Du8Chuanxin Wang9Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, ChinaDepartment of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, ChinaUsually, 18–48 h are needed for the identification of microbial pathogens causing urinary tract infections (UTIs) by urine culture. Moreover, antimicrobial susceptibility testing (AST) takes an additional 18–24 h. Rapid identification and AST of the pathogens allow fast and precise treatment. The objective of this study was to shorten the time of diagnosis of UTIs by combining pathogen screening through flow cytometry, microbial identification by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS), and AST using the VITEK 2 system for the direct analysis of urine samples. We analyzed 1,638 urine samples from patients with suspected UTIs submitted to the microbiology laboratory for culture. Each urine sample had an approximate volume of 30 mL and was divided into three aliquots. Urine processing included differential centrifugation and two washes to enrich the bacterial fraction for direct MALDI-TOF MS and direct AST. From a total of 1,638 urine samples, 307 were found to be positive through UF-1000i screening. Among them, 265 had significant growth of a single-microorganism. Direct identification was obtained in 229 (86.42%) out of these 265 samples, and no pathogens were misidentified. Moreover, species-level identification was obtained in 163 (88.59%) out of the 184 samples with Gram-negative bacteria, and 27 (38.03%) out of the 71 samples with Gram-positive bacteria. VITEK 2 AST was performed for 117 samples with a single-microorganism. Enterobacteriaceae data showed an agreement rate of antimicrobial categories of 94.83% (1,229/1,296), with minor, major, and very major error rates of 4.17% (54/1,296), 0.92% (12/1,296), and 0.08% (1/1,296), respectively. For Enterococcus spp., the overall categorical agreement was 92.94% (158/170), with a minor error rate of 2.94% (5/170) and major error rate of 4.12% (7/170). The turnaround time of this combined protocol to diagnose UTIs was 1 h for pathogen identification and 6–24 h for AST; noteworthily, only 6–8 h are needed for AST of Enterobacteriaceae using the VITEK 2 system. Overall, our findings show that the combination of flow cytometry, MALDI-TOF MS, and VITEK 2 provided a direct, rapid, and reliable identification and AST method for assessing urine samples, especially for Gram-negative bacterial infections.https://www.frontiersin.org/article/10.3389/fmicb.2019.01182/fullbacterial identificationantimicrobial susceptibilityMALDI-TOF MSurinary tract infectionsrapid diagnosisurine |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Wei Li Enhua Sun Ying Wang Hongwei Pan Yi Zhang Yong Li Xin Zhang Chen Li Lutao Du Chuanxin Wang |
spellingShingle |
Wei Li Enhua Sun Ying Wang Hongwei Pan Yi Zhang Yong Li Xin Zhang Chen Li Lutao Du Chuanxin Wang Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol Frontiers in Microbiology bacterial identification antimicrobial susceptibility MALDI-TOF MS urinary tract infections rapid diagnosis urine |
author_facet |
Wei Li Enhua Sun Ying Wang Hongwei Pan Yi Zhang Yong Li Xin Zhang Chen Li Lutao Du Chuanxin Wang |
author_sort |
Wei Li |
title |
Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol |
title_short |
Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol |
title_full |
Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol |
title_fullStr |
Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol |
title_full_unstemmed |
Rapid Identification and Antimicrobial Susceptibility Testing for Urinary Tract Pathogens by Direct Analysis of Urine Samples Using a MALDI-TOF MS-Based Combined Protocol |
title_sort |
rapid identification and antimicrobial susceptibility testing for urinary tract pathogens by direct analysis of urine samples using a maldi-tof ms-based combined protocol |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Microbiology |
issn |
1664-302X |
publishDate |
2019-06-01 |
description |
Usually, 18–48 h are needed for the identification of microbial pathogens causing urinary tract infections (UTIs) by urine culture. Moreover, antimicrobial susceptibility testing (AST) takes an additional 18–24 h. Rapid identification and AST of the pathogens allow fast and precise treatment. The objective of this study was to shorten the time of diagnosis of UTIs by combining pathogen screening through flow cytometry, microbial identification by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS), and AST using the VITEK 2 system for the direct analysis of urine samples. We analyzed 1,638 urine samples from patients with suspected UTIs submitted to the microbiology laboratory for culture. Each urine sample had an approximate volume of 30 mL and was divided into three aliquots. Urine processing included differential centrifugation and two washes to enrich the bacterial fraction for direct MALDI-TOF MS and direct AST. From a total of 1,638 urine samples, 307 were found to be positive through UF-1000i screening. Among them, 265 had significant growth of a single-microorganism. Direct identification was obtained in 229 (86.42%) out of these 265 samples, and no pathogens were misidentified. Moreover, species-level identification was obtained in 163 (88.59%) out of the 184 samples with Gram-negative bacteria, and 27 (38.03%) out of the 71 samples with Gram-positive bacteria. VITEK 2 AST was performed for 117 samples with a single-microorganism. Enterobacteriaceae data showed an agreement rate of antimicrobial categories of 94.83% (1,229/1,296), with minor, major, and very major error rates of 4.17% (54/1,296), 0.92% (12/1,296), and 0.08% (1/1,296), respectively. For Enterococcus spp., the overall categorical agreement was 92.94% (158/170), with a minor error rate of 2.94% (5/170) and major error rate of 4.12% (7/170). The turnaround time of this combined protocol to diagnose UTIs was 1 h for pathogen identification and 6–24 h for AST; noteworthily, only 6–8 h are needed for AST of Enterobacteriaceae using the VITEK 2 system. Overall, our findings show that the combination of flow cytometry, MALDI-TOF MS, and VITEK 2 provided a direct, rapid, and reliable identification and AST method for assessing urine samples, especially for Gram-negative bacterial infections. |
topic |
bacterial identification antimicrobial susceptibility MALDI-TOF MS urinary tract infections rapid diagnosis urine |
url |
https://www.frontiersin.org/article/10.3389/fmicb.2019.01182/full |
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