ORIGINAL RESEARCH article
Front. Cell. Infect. Microbiol.
Sec. Clinical and Diagnostic Microbiology and Immunology
A MALDI-TOF nucleic acid mass spectrometry assay for rapid detection of five Candida species in urine
Provisionally accepted
- 1The Second Affiliated Hospital of Xuzhou Medical University, Xuzhou, China
- 2Chongqing General Hospital, Chongqing, China
- 3The Affiliated Hospital of Xuzhou Medical University, Xuzhou, China
- 4Xuzhou Medical University, Xuzhou, China
- 5Affiliated Hospital of Xuzhou Medical University, Xuzhou, China
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Background: Repeat urine testing is key for the management of candiduria. Current urine testing methods are time-consuming. This study presents an assay coupling PCR with matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS), termed MALDI-TOF nucleic acid MS (MALDI-TOF NAMS), for rapid detection of five medically important Candida species (Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis and Candida krusei ) in urine. Methods: Species-specific primers and probes targeting the internal transcribed spacer (ITS) regions the five Candida species were designed, followed by PCR amplification, single-base extension, and MALDI-TOF MS analysis. The sensitivity assessment was conducted using Candida suspensions and simulated candiduria samples. DNA from bacteria and other fungi was tested to analyse the specificity. To evaluate multiplex detection capability, all five Candida species were simultaneously tested in a single-tube reaction. Clinical performance was validated using collected urine samples, with results compared with urine culture. Results The developed assay demonstrated a limit of detection (LoD) ranging from 101 CFU/mL to 103 CFU/mL across different Candida species, with no cross-reactivity against common uropathogens. The assay allowed simultaneous detection of multiple Candida species in a single tube. The diagnostic sensitivity of our developed method was 100% using urine culture as the gold standard, with a diagnostic specificity of 98.7%. The entire detection process could be completed within 5 hours. Conclusion The novel MALDI-TOF NAMS assay enables rapid detection of five Candida species in urine with validated performance. The assay supports clinical candiduria management and is adaptable for broader Candida species detection.
Keywords: Candida, Urine, MALDI-TOF NAMS, ITS, Candiduria
Received: 09 Aug 2025; Accepted: 24 Oct 2025.
Copyright: © 2025 Guo, Chen, Sun, Zhu, Haiquan and Xu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Kang Haiquan, hqk811029@163.com
Yinhai Xu, dolmguoyi@xzhmu.edu.cn
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