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.
PLoS One. 2025 Aug 20;20(8):e0330455. doi: 10.1371/journal.pone.0330455. eCollection 2025.
Fungal keratitis (FK) is a sight-threatening infectious disease that can result in blindness if not appropriately treated. Although keratitis associated with filamentous fungi was rarely reported in the past, the incidence of fungal keratitis due to contact lens usage has increased. The clinical manifestations of fungal keratitis often resemble those of other corneal infections, potentially delaying accurate diagnosis and treatment. In this study, we developed a Fusarium-specific polyclonal peptide antibody targeting the cutinase of F. solani and evaluated its diagnostic potential for FK in an animal model. To assess the specificity of the cutinase antibody, we employed enzyme-linked immunosorbent assay (ELISA). The ELISA results demonstrated that the cutinase antibody specifically interacted with the cell lysates and conditioned media of F. solani. Additionally, an immunocytochemistry assay confirmed the specificity of the cutinase antibody for F. solani in samples co-cultured with human corneal epithelial (HCE) cells and other keratitis-causing agents. To validate our in vitro findings, FK animal models were established by infecting the corneas of BALB/c mice with F. solani. The cutinase antibody specifically detected Fusarium antigens in the tear-wash samples and eyeball lysates of FK mice. These results demonstrate that the cutinase antibody is highly specific to F. solani antigens, indicating its potential utility in developing an antibody-based diagnostic method for FK.
真菌性角膜炎（FK）是一种威胁视力的感染性疾病，若未得到恰当治疗可导致失明。尽管过去与丝状真菌相关的角膜炎鲜有报道，但因佩戴隐形眼镜导致的真菌性角膜炎发病率有所上升。真菌性角膜炎的临床表现常与其他角膜感染相似，可能会延误准确诊断和治疗。在本研究中，我们开发了一种针对茄病镰刀菌角质酶的镰刀菌特异性多克隆肽抗体，并在动物模型中评估了其对真菌性角膜炎的诊断潜力。为评估角质酶抗体的特异性，我们采用了酶联免疫吸附测定（ELISA）。ELISA结果表明，角质酶抗体与茄病镰刀菌的细胞裂解物和条件培养基发生特异性相互作用。此外，免疫细胞化学分析证实了角质酶抗体在与人角膜上皮（HCE）细胞及其他致角膜炎病原体共培养的样本中对茄病镰刀菌的特异性。为验证我们的体外研究结果，通过用茄病镰刀菌感染BALB/c小鼠的角膜建立了真菌性角膜炎动物模型。角质酶抗体在真菌性角膜炎小鼠的泪液冲洗样本和眼球裂解物中特异性检测到了镰刀菌抗原。这些结果表明，角质酶抗体对茄病镰刀菌抗原具有高度特异性，表明其在开发基于抗体的真菌性角膜炎诊断方法方面具有潜在用途。
