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Rapid Identification of Four Fusarium spp. Complex by High-Resolution Melting Curve Analysis and their Antifungal Susceptibility Profiles

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Abstract

Fusarium species are globally distributed filamentous ascomycete fungi that are frequently reported as plant pathogens and opportunistic human pathogens, leading to yield loss of crops, mycotoxin contamination of food and feed products as well as damage to human and livestock. Human infections of Fusarium spp. are difficult to treat due to broad antifungal resistance by members of this genus. Their role as disease-causing agents in crops and humans suggests a need for antifungal resistance profiles as well as a simple, rapid, and cost effective identification method. Fusarium strains were isolated from food and clinical samples. High-resolution melting curve (HRM) analysis was performed using specific primers targeting internal transcribed spacer (ITS) region, followed with evaluation of specificity and sensitivity. The antifungal susceptibility of four Fusarium species was studied using the Sensititre YeastOne method. HRM analysis revealed reproducible, unimodal melting profiles specific to each of the four Fusarium strains, while no amplification of the negative controls. The minimum detection limits were 100–120 copies based on a 2 µl volume of template. Clear susceptibility differences were observed against antifungal agents by different Fusarium isolates, with amphotericin B and voriconazole displayed strongest antifungal effects to all the tested strains. We developed a simple, rapid, and low-cost qPCR-HRM method for identification of four Fusarium spp. (F. oxysporum, F. lateritium, F. fujikuroi, and F. solani). The antifungal susceptibility profiles supplied antifungal information of foodborne and clinical Fusarium spp. and provided guidance for clinical treatment of human infections.

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Funding

This work was supported by National Key Research and Development Program of China (Grant Number 2021YFC2401000), National Key Research and Development Program of China (Grant Number 2018YFC1603801) and National Science and Technology Major Project of China (Grant Number 2018ZX10712001-006-007).

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Authors and Affiliations

  1. State Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changbai Road 155, Changping, Beijing, 102206, China

    Xuexin Hou, Yuanyuan Geng, Fei Zhao, Lihua He & Jie Gong

  2. College of Public Health, Zhejiang Chinese Medical University, No. 548, Binwen Road, Binjiang District, Hangzhou, 310053, Zhejiang, China

    Rongchen Dai

Authors
  1. Xuexin Hou
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  2. Yuanyuan Geng
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  3. Rongchen Dai
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  4. Fei Zhao
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  5. Lihua He
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  6. Jie Gong
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Contributions

Resources, data curation and the first draft of the manuscript were performed by Xuexin Hou; methodology, data collection and analysis were performed by Yuanyuan Geng, Rongchen Dai, Fei Zhao and Lihua He contributed to the software, investigation and project administration. Conceptualization, supervision and funding acquisition were performed by Jie Gong. All authors read and approved the final manuscript.

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Correspondence to Jie Gong.

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The authors have no relevant financial or non-financial interests to disclose.

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Handling Editor: Weida Liu.

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X. Hou and Y. Geng contribute equally to this work.

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Supporting Information

Supplemental Data 1: Concentrations of each antifungal drug tested in this study.

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Hou, X., Geng, Y., Dai, R. et al. Rapid Identification of Four Fusarium spp. Complex by High-Resolution Melting Curve Analysis and their Antifungal Susceptibility Profiles. Mycopathologia 187, 345–354 (2022). https://doi.org/10.1007/s11046-022-00635-8

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  • DOI: https://doi.org/10.1007/s11046-022-00635-8

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