收稿日期: 2025-05-08
网络出版日期: 2025-08-28
基金资助
国家自然科学基金青年科学基金(82402722);徐州医科大学附属医院院级科研项目(2021ZA25)
Application value of nucleic acid mass spectrometry in detecting ERG11 mutation in Candida tropicalis
Received date: 2025-05-08
Online published: 2025-08-28
目的 探讨核酸质谱技术在热带念珠菌唑类耐药相关基因ERG11突变位点检测中的应用价值，为临床唑类药物的合理规范使用提供可靠依据。 方法 收集徐州医科大学附属医院2023年7月至2024年11月分离的热带念珠菌菌株，对同一菌株进行Sanger测序和核酸质谱检测ERG11基因A395T和C461T的突变情况。以Sanger测序结果为金标准，评价核酸质谱技术检测位点突变的性能。 结果 本研究所构建的核酸质谱检测方法可同时检测出ERG11的 A395T和C461T的4种基因型，各突变位点均可产生特异产物峰且无相互干扰。在88株热带念珠菌临床分离株中，A395T位点突变的敏感度、特异度、阳性预测值、阴性预测值分别为86.7%、100%、100%、97.3%；C461T位点突变的敏感度、特异度、阳性预测值、阴性预测值分别为81.3%、100%、100%、96.0%。Kappa检验显示核酸质谱技术与测序方法一致性强。 结论 核酸质谱技术检测ERG11位点突变灵敏度、特异度均较高，具有快速、灵敏、准确、通量灵活的特性，是真菌耐药基因突变位点检测的有效方法。
陈燕 , 朱红霞 , 康海全 , 郭毅 , 徐银海 , 孙静芳 . 核酸质谱技术在热带念珠菌ERG11位点突变检测中的应用[J]. 实用医学杂志, 2025 , 41(16) : 2575 -2580 . DOI: 10.3969/j.issn.1006-5725.2025.16.020
Objective To evaluate the application value of nucleic acid mass spectrometry in detecting ERG11 gene mutations associated with azole resistance in Candida tropicalis （C. tropicalis）， thereby providing a scientific basis for the rational clinical use of azole antifungal agents. Methods Clinical isolates of C. tropicalis were obtained from the Affiliated Hospital of Xuzhou Medical University between July 2023 and November 2024. For each isolate， specific ERG11 mutations （A395T and C461T） were analyzed using Sanger sequencing and nucleic acid mass spectrometry. Sanger sequencing was used as the reference standard to evaluate the performance of mass spectrometry in mutation detection. Results The established nucleic acid mass spectrometry method effectively identified four genotypes of ERG11 （A395T and C461T）， with distinct spectral peaks for each mutation and no cross-interference observed. Among 88 clinical isolates， the sensitivity， specificity， positive predictive value （PPV）， and negative predictive value （NPV） for the A395T mutation were 86.7%， 100%， 100%， and 97.3%， respectively. For the C461T mutation， the corresponding values were 81.3%， 100%， 100%， and 96.0%， respectively. Kappa statistics demonstrated a high level of agreement between mass spectrometry and sequencing results. Conclusions Nucleic acid mass spectrometry exhibits high sensitivity and specificity in the detection of ERG11 mutations， enabling rapid， accurate， and adaptable high-throughput analysis. This makes it a highly effective method for identifying mutations associated with fungal resistance.
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