眼科 ›› 2026, Vol. 35 ›› Issue (1): 27-32.doi: 10.13281/j.cnki.issn.1004-4469.2026.01.004

• 论著 • 上一篇    下一篇

角膜刮片细胞学检查不同染色方法和宏基因组二代测序在眼部微孢子虫感染诊断中的应用

张阳1 梁庆丰1 王智群1 何彦2 徐英男3 屈虹男4 陈可心1 邓世靖2 韩冰4   

  1. 1首都医科大学附属北京同仁医院 北京同仁眼科中心 北京市眼科研究所 眼科学与视觉科学北京市重点实验室,北京 100730;2首都医科大学附属北京同仁医院 北京同仁眼科中心 眼科学与视觉科学北京市重点实验室,北京 100730;3南京医科大学附属眼科医院,南京 210029;4山东大学基础医学院病原生物学系,济南250012
  • 收稿日期:2025-11-12 出版日期:2026-01-25 发布日期:2026-01-13
  • 通讯作者: 韩冰,Email: bing.han@sdu.edu.cn E-mail:bing.han@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(82572601,82502750);南京市卫生科技发展基金(ZDXX25212)

Application of different staining methods in corneal scraping cytology and metagenomic next-generation sequencing in diagnosis of ocular microsporidiosis

Zhang Yang1, Liang Qingfeng1, Wang Zhiqun1, He Yan2, Xu Yingnan3, Qu Hongnan4, Chen Kexin1, Deng Shijing2, Han Bing4   

  1. 1 Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University; Beijing Key Laboratory of Ophthalmology and Visual Sciences, Beijing 100730, China; 2 Beijing Tongren Eye Center, Capital Medical University, Beijing Tongren Hospital; Beijing Key Laboratory of Ophthalmology and Visual Sciences, Beijing 100730, China; 3 Eye Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China; 4 Department of Pathogenic Biology, School of Basic Medical Sciences, Shandong University, Jinan Shandong 250012, China
  • Received:2025-11-12 Online:2026-01-25 Published:2026-01-13
  • Contact: Han Bing, Email: bing.han@sdu.edu.cn E-mail:bing.han@sdu.edu.cn
  • Supported by:
    National Natural Science Foundation of China (82572601, 82502750); Nanjing Health Science and Technology Development Fund (ZDXX25212)

摘要: 目的 探讨角膜刮片细胞学检查中不同染色方法与宏基因组二代测序对眼部微孢子虫感染的诊断价值。设计 回顾性病例系列。研究对象 北京同仁医院眼科接诊的 33 例(40 眼)眼部微孢子虫感染患者。方法 采用四种染色法(吉姆萨染色,真菌荧光染色,革兰染色,抗酸染色)行角膜刮片细胞学检查,结合宏基因组二代测序(metagenomic next-generation sequencing,mNGS),分析不同方法检出率及mNGS病原谱。主要指标 不同染色方法的检出率。结果 刮片细胞学整体检出率 90.9%,其中吉姆萨染色、真菌荧光染色、革兰染色、抗酸染色的阳性检出率分别为 81.8%、63.6%、24.2%、9.1%。吉姆萨联合真菌荧光染色检出率为90.9%。采用Pearson 卡方检验分析发现四种染色方法的阳性检出率存在显著差异(χ2=33.056, df=3, P<0.001)。mNGS 检出率为 93.9%,检测出 5 种微孢子虫。结论 角膜刮片细胞学检查与宏基因组二代测序两种方法均为有效诊断手段,推荐吉姆萨染色联合真菌荧光染色。

关键词: 微孢子虫, 微孢子虫性角结膜炎, 角膜刮片, 宏基因组二代测序

Abstract: Objective To explore the diagnostic value of different corneal scraping cytology staining methods and metagenomic next-generation sequencing (mNGS) for ocular microsporidiosis. Design Retrospective study. Participants Thirty-three patients (40 eyes) with ocular microsporidiosis admitted to Beijing Tongren Hospital. Methods Corneal scraping cytology was performed with four staining methods (Giemsa, fungal fluorescence, Gram, acid-fast staining). Combined with mNGS, the detection rates of each method and the pathogenic spectrum of mNGS were analyzed. Main Outcome Measures Detection rates of different staining methods and mNGS. Results The overall detection rate of scraping cytology was 90.9%. The detection rates of Giemsa, fungal fluorescence, Gram, and acid-fast staining were 81.8%, 63.6%, 24.2%, and 9.1%, respectively. The combined Giemsa and fungal fluorescence staining achieved the highest detection rate (90.9%). Pearson's chi-square test showed significant differences in positive rates among the four staining methods (χ2=33.056, df=3, P<0.001). The detection rate of mNGS was 93.9%, with 5 microsporidia species identified. Conclusion Both corneal scraping cytology and mNGS are effective for diagnosing ocular microsporidiosis. Combination of Giemsa and fungal fluorescence staining is recommended.

Key words: Microsporidia, Microsporidial keratoconjunctivitis, Corneal scraping, Metagenomic next-generation sequencing