Jo Hye-Jeong, Kim Min-Jeong, Lee Hae-Ahm, Quan Fu-Shi, Kong Hyun-Hee, Moon Eun-Kyung
Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul, Republic of Korea.
Department of Parasitology, School of Medicine, Chungbuk National University, Cheongju, Republic of Korea.
Sci Rep. 2025 Jul 1;15(1):21583. doi: 10.1038/s41598-025-08719-3.
The increasing number of contact lens users correlates with a rise in the incidence of fungal keratitis. Fungal keratitis can lead to blindness if not treated promptly, and its early symptoms are similar to those of bacterial and amoebic keratitis, making rapid diagnosis challenging. This study aimed to assess the potential of using a peptide antibody against the fungal-specific protein ERG24, which encodes sterol C-14 reductase, to differentiate fungal keratitis from other forms of keratitis. The specificity of the ERG24 antibody was assessed through Western blot and enzyme-linked immunosorbent assay (ELISA). Immunocytochemistry (ICC) was performed by co-culturing two types of fungi, Acanthamoeba, and two bacterial strains with human corneal epithelial (HCE) cells. Additionally, to evaluate the diagnostic potential of the ERG24 antibody, animal models of fungal, amoebic, and bacterial keratitis were developed, and ELISA was conducted on tear and ocular lysates from these models. The results demonstrated that the antibody specifically reacted with Fusarium solani in Western blot, and both ELISA and ICC confirmed that the ERG24 antibody did not react with HCE cells, Acanthamoeba, or bacteria, but was specific to the two fungal species. In vivo experiments further showed that the ERG24 antibody significantly detected F. solani in tear-wash samples and eyeball lysates from the fungal keratitis model, without reacting with samples from amoebic and bacterial keratitis models. This study suggests that the ERG24 peptide antibody could provide valuable information for developing differential diagnostic methods for fungal keratitis compared to other forms of keratitis.
隐形眼镜使用者数量的增加与真菌性角膜炎发病率的上升相关。真菌性角膜炎若不及时治疗可导致失明，其早期症状与细菌性和阿米巴性角膜炎相似，使得快速诊断具有挑战性。本研究旨在评估使用针对真菌特异性蛋白ERG24（编码甾醇C - 14还原酶）的肽抗体来区分真菌性角膜炎与其他形式角膜炎的潜力。通过蛋白质免疫印迹法和酶联免疫吸附测定（ELISA）评估ERG24抗体的特异性。通过将两种真菌、阿米巴原虫和两种细菌菌株与人角膜上皮（HCE）细胞共培养来进行免疫细胞化学（ICC）。此外，为了评估ERG24抗体的诊断潜力，建立了真菌性、阿米巴性和细菌性角膜炎的动物模型，并对这些模型的泪液和眼组织裂解液进行ELISA检测。结果表明，该抗体在蛋白质免疫印迹中与茄病镰刀菌特异性反应，ELISA和ICC均证实ERG24抗体不与HCE细胞、阿米巴原虫或细菌反应，而是对这两种真菌具有特异性。体内实验进一步表明，ERG24抗体在真菌性角膜炎模型的泪液冲洗样本和眼球裂解液中能显著检测到茄病镰刀菌，而不与阿米巴性和细菌性角膜炎模型的样本反应。本研究表明，与其他形式的角膜炎相比，ERG24肽抗体可为开发真菌性角膜炎的鉴别诊断方法提供有价值的信息。
