Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification
IntroductionCandida albicans, a human fungal pathogen, multiplies to invade body cells and causes fungal diseases in the condition of insufficient body's immune function. Early detection of C. albicans is required to guide appropriate prevention and treatment.MethodsThe purpose of this study wa...
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Frontiers Media S.A.
2025-02-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1484089/full |
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| author | Yanli Zhang Xuhan Chen Yeling Zhong Fei Guo Guifang Ouyang Rui Mao |
| author_facet | Yanli Zhang Xuhan Chen Yeling Zhong Fei Guo Guifang Ouyang Rui Mao |
| author_sort | Yanli Zhang |
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| description | IntroductionCandida albicans, a human fungal pathogen, multiplies to invade body cells and causes fungal diseases in the condition of insufficient body's immune function. Early detection of C. albicans is required to guide appropriate prevention and treatment.MethodsThe purpose of this study was to establish a C. albicans assay based on newly developed closed dumbbell-mediated isothermal amplification (CDA) to achieve rapid and simple point of care diagnostic. The CDA technique was carried out by specific primers targeting at the conserved C. albicans ITS2 gene. All primers were selected and evaluated by real-time fluorescence monitoring and endpoint visual judgement indicated by hydroxy naphthol blue (HNB). Optimal primers and accelerate primers (out primers and loop primers) were designed and selected after confirmation of the fundamental CDA primers to achieve more efficient CDA reaction for C. albicans detection (CA-OL-CDA).ResultsAfter establishment of the assay, 9 non-Candida albicans strains, including 3 Candida species were tested to negative by adopting the established CA-OL-CDA assay, indicated high specificity. The limit of detection of Candida albicans DNA by CA-OL-CDA assay was 6.2×10-6 ng/μL of DNA (10 copies/μL), 10-fold more sensitive than real-time quantitative PCR (qPCR).DiscussionThe CA-OL-CDA assay exhibited advantages of high specificity, sensitivity, simpler and more efficient operation. In addition, the CA-OL-CDA method holds potential in on-site detection for C. albicans using color shift by adopting the reaction mixture based on HNB. |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
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| spelling | doaj-art-e9fa5571d8fe4dc3b0735deea1552cf62025-02-03T06:33:19ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-02-011510.3389/fcimb.2025.14840891484089Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplificationYanli Zhang0Xuhan Chen1Yeling Zhong2Fei Guo3Guifang Ouyang4Rui Mao5Department of Hematology, The First Affiliated Hospital of Ningbo University, Ningbo, Zhejiang, ChinaNingbo Institute of Life and Health Industry, University of Chinese Academy of Sciences, Ningbo, ChinaDepartment of General Surgery (Hepatic, Anal-canal, Gastrointestinal), Ningbo Zhenhai People’s Hospital, Ningbo, Zhejiang, ChinaDepartment of Laboratory Medicine, The First Affiliated Hospital of Ningbo University, Ningbo, Zhejiang, ChinaDepartment of Hematology, The First Affiliated Hospital of Ningbo University, Ningbo, Zhejiang, ChinaNingbo Institute of Life and Health Industry, University of Chinese Academy of Sciences, Ningbo, ChinaIntroductionCandida albicans, a human fungal pathogen, multiplies to invade body cells and causes fungal diseases in the condition of insufficient body's immune function. Early detection of C. albicans is required to guide appropriate prevention and treatment.MethodsThe purpose of this study was to establish a C. albicans assay based on newly developed closed dumbbell-mediated isothermal amplification (CDA) to achieve rapid and simple point of care diagnostic. The CDA technique was carried out by specific primers targeting at the conserved C. albicans ITS2 gene. All primers were selected and evaluated by real-time fluorescence monitoring and endpoint visual judgement indicated by hydroxy naphthol blue (HNB). Optimal primers and accelerate primers (out primers and loop primers) were designed and selected after confirmation of the fundamental CDA primers to achieve more efficient CDA reaction for C. albicans detection (CA-OL-CDA).ResultsAfter establishment of the assay, 9 non-Candida albicans strains, including 3 Candida species were tested to negative by adopting the established CA-OL-CDA assay, indicated high specificity. The limit of detection of Candida albicans DNA by CA-OL-CDA assay was 6.2×10-6 ng/μL of DNA (10 copies/μL), 10-fold more sensitive than real-time quantitative PCR (qPCR).DiscussionThe CA-OL-CDA assay exhibited advantages of high specificity, sensitivity, simpler and more efficient operation. In addition, the CA-OL-CDA method holds potential in on-site detection for C. albicans using color shift by adopting the reaction mixture based on HNB.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1484089/fullCandida albicanspoint-of-care diagnosticclosed dumbbell mediated isothermal amplificationreal-time fluorescence CDAvisual CDAsensitivity |
| spellingShingle | Yanli Zhang Xuhan Chen Yeling Zhong Fei Guo Guifang Ouyang Rui Mao Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification Frontiers in Cellular and Infection Microbiology Candida albicans point-of-care diagnostic closed dumbbell mediated isothermal amplification real-time fluorescence CDA visual CDA sensitivity |
| title | Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification |
| title_full | Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification |
| title_fullStr | Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification |
| title_full_unstemmed | Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification |
| title_short | Rapid and simple detection of Candida albicans using closed dumbbell-mediated isothermal amplification |
| title_sort | rapid and simple detection of candida albicans using closed dumbbell mediated isothermal amplification |
| topic | Candida albicans point-of-care diagnostic closed dumbbell mediated isothermal amplification real-time fluorescence CDA visual CDA sensitivity |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1484089/full |
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