Development and evaluation of a centrifugal disk system for the rapid detection of multiple pathogens and their antibiotic resistance genes in urinary tract infection

Frontiers in Microbiology

Microbiology

Ming Zong¹  Wenyan Zhang¹  Nianzhen Chen¹  Yuying Si¹  Gen Li¹  Yangqin Ye¹  Lieying Fan¹  Tong Zhang²  Lifeng Pan³  Dali Chi⁴  Yun Lu⁵  Guodong Sui⁶  Guoying Qu⁷ 

[1] Department of Clinical Laboratory, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China;Department of Clinical Laboratory, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China;Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention (LAP3), Department of Environmental Science and Engineering, Fudan University, Shanghai, China;Department of Microbiology, Pudong New Area Center for Disease Control and Prevention, Shanghai, China;Fosun Diagnostics Co., Ltd., Shanghai, China;Huamu Community Health Service Center, Shanghai, China;Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention (LAP3), Department of Environmental Science and Engineering, Fudan University, Shanghai, China;Weifang Community Health Service Center, Shanghai, China;

关键词: LAMP;    centrifugal disk;    urinary tract infection;    pathogens;    antibiotic resistance genes;   

DOI  :  10.3389/fmicb.2023.1157403

 received in 2023-02-02, accepted in 2023-03-08,  发布年份 2023

来源: Frontiers

【 摘 要 】

BackgroundUrinary tract infections (UTIs) are some of the most common bacterial infections in the world. Nevertheless, as uncomplicated UTIs are treated empirically without culturing the urine, adequate knowledge of the resistance pattern of uropathogens is essential. Conventional urine culture and identification take at least 2 days. Here, we developed a platform based on LAMP and centrifugal disk system (LCD) to simultaneously detect the main pathogens and antibiotic resistant genes (ARGs) of urgent concern multidrug-resistant among UTIs.MethodsWe designed specific primers to detect the target genes above and evaluated their sensitivity and specificity. We also assessed the result of our preload LCD platform on 645 urine specimens with a conventional culturing method and Sanger sequencing.ResultsThe results obtained with the 645 clinical samples indicated that the platform has high specificity (0.988–1) and sensitivity (0.904–1) for the studied pathogens and ARGs. Moreover, the kappa value of all pathogens was more than 0.75, revealing an excellent agreement between the LCD and culture method. Compared to phenotypic tests, the LCD platform is a practical and fast detection approach for methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococci, carbapenem-resistant Enterobacteriaceae, carbapenem-resistant Acinetobacter, carbapenem-resistant Pseudomonas aeruginosa (kappa value of all >0.75), and non-extended-spectrum β-lactamase producers.ConclusionWe developed a detection platform that has high accuracy and that meets the need for rapid diagnosis, which can be completed within 1.5 h from specimen collection. It may be a powerful tool for evidence-based UTIs diagnosis, which has essential support for the rational use of antibiotics. More high-quality clinical studies are required to prove the effectiveness of our platform.

【 授权许可】

Unknown   
Copyright © 2023 Chen, Li, Si, Ye, Zhang, Chi, Zhang, Pan, Qu, Lu, Zong, Sui and Fan.