Kim Min-Jeong, Jo Hye-Jeong, Sohn Hae-Jin, Shin Ho-Joon, Quan Fu-Shi, Kong Hyun-Hee, Moon Eun-Kyung
Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea.
Department of Microbiology, Ajou University School of Medicine, Suwon 16499, Republic of Korea.
Pathogens. 2023 Mar 28;12(4):526. doi: 10.3390/pathogens12040526.
spp. is the causative agent of keratitis (AK), a vision-threatening parasitic disease whose primary risk factor has been attributed to poor contact lens hygiene. Unfortunately, differential diagnosis of AK is challenging as the clinical manifestations for AK are similar to those of bacterial, fungal, or even viral keratitis. Since delayed AK diagnosis can incur permanent vision impairment, a rapid and sensitive diagnostic method is urgently needed. Here, the diagnostic potential of polyclonal antibodies targeting the chorismate mutase (CM) of spp. was evaluated in AK animal models. CM antibody specificity against trophozoites and cysts was confirmed by immunocytochemistry after co-culturing with , , and , and human corneal epithelial (HCE) cells. Enzyme-linked immunosorbent assay (ELISA) was performed using CM-specific immune sera raised in rabbits, which demonstrated that the antibodies specifically interacted with the trophozoites and cysts in a dose-dependent manner. To evaluate the diagnostic potential of the CM antibody, AK animal models were established by incubating contact lenses with an inoculum containing trophozoites and subsequently overlaying these lenses onto the corneas of BALB/c mice for 7 and 21 days. The CM antibody specifically detected antigens in the murine lacrimal and eyeball tissue lysates at both time points. Our findings underscore the importance of antibody-based AK diagnosis, which could enable early and differential AK diagnosis in clinical settings.
某属物种是角膜溃疡（AK）的病原体，这是一种威胁视力的寄生虫病，其主要风险因素被认为是隐形眼镜卫生状况不佳。不幸的是，AK的鉴别诊断具有挑战性，因为AK的临床表现与细菌性、真菌性甚至病毒性角膜炎相似。由于AK诊断延迟会导致永久性视力损害，因此迫切需要一种快速且灵敏的诊断方法。在此，在AK动物模型中评估了针对某属物种分支酸变位酶（CM）的多克隆抗体的诊断潜力。在与某属物种、其他物种以及人角膜上皮（HCE）细胞共培养后，通过免疫细胞化学证实了CM抗体对某属物种滋养体和包囊的特异性。使用在兔体内产生的CM特异性免疫血清进行酶联免疫吸附测定（ELISA），结果表明抗体以剂量依赖方式与某属物种滋养体和包囊特异性相互作用。为了评估CM抗体的诊断潜力，通过将含有某属物种滋养体的接种物与隐形眼镜孵育，随后将这些隐形眼镜覆盖在BALB/c小鼠的角膜上7天和21天，建立了AK动物模型。在这两个时间点，CM抗体均能特异性检测到小鼠泪液和眼球组织裂解物中的某属物种抗原。我们的研究结果强调了基于抗体的AK诊断的重要性，这能够在临床环境中实现AK的早期和鉴别诊断。
