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

Previous Articles     Next Articles

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)

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