Li Wei, Sun Enhua, Wang Ying, Pan Hongwei, Zhang Yi, Li Yong, Zhang Xin, Li Chen, Du Lutao, Wang Chuanxin
Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, China.
Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, China.
Front Microbiol. 2019 Jun 5;10:1182. doi: 10.3389/fmicb.2019.01182. eCollection 2019.
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. 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 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 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.
通常，通过尿培养鉴定引起尿路感染（UTIs）的微生物病原体需要18 - 48小时。此外，抗菌药物敏感性测试（AST）还需要额外的18 - 24小时。病原体的快速鉴定和AST能够实现快速且精确的治疗。本研究的目的是通过结合流式细胞术进行病原体筛查、基质辅助激光解吸电离飞行时间质谱（MALDI - TOF MS）进行微生物鉴定以及使用VITEK 2系统对尿液样本进行直接分析的AST，来缩短UTIs的诊断时间。我们分析了提交至微生物实验室进行培养的1638例疑似UTIs患者的尿液样本。每个尿液样本的体积约为30 mL，并分为三等份。尿液处理包括差速离心和两次洗涤，以富集细菌部分用于直接MALDI - TOF MS和直接AST。在总共1638份尿液样本中，通过UF - 1000i筛查发现307份为阳性。其中，265份有单一微生物的显著生长。在这265份样本中，229份（86.42%）获得了直接鉴定，且没有病原体被错误鉴定。此外，在184份革兰氏阴性菌样本中的163份（88.59%）以及71份革兰氏阳性菌样本中的27份（38.03%）获得了种水平鉴定。对117份单一微生物样本进行了VITEK 2 AST。数据显示抗菌类别一致率为94.83%（1229/1296），轻微、主要和非常主要错误率分别为4.17%（54/1296）、0.92%（12/1296）和0.08%（1/1296）。对于 spp.，总体类别一致率为92.94%（158/170），轻微错误率为2.94%（5/170），主要错误率为4.12%（7/170）。这种联合方案诊断UTIs的周转时间为病原体鉴定1小时，AST为6 - 24小时；值得注意的是，使用VITEK 2系统对 进行AST仅需6 - 8小时。总体而言，我们的研究结果表明，流式细胞术、MALDI - TOF MS和VITEK 2的联合为评估尿液样本，特别是革兰氏阴性菌感染，提供了一种直接、快速且可靠的鉴定和AST方法。
Clin Microbiol Infect. 2016-2-17
Antibiotics (Basel). 2022-5-15
Acta Microbiol Immunol Hung. 2020-9-24
Eur J Clin Microbiol Infect Dis. 2024-11
Microorganisms. 2023-5-16
Nat Rev Urol. 2017-3-1
Curr Opin Urol. 2017-3
Trends Mol Med. 2016-11
Zotero 插件