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Detection of Acanthamoeba spp. using carboxylesterase antibody and its usage for diagnosing Acanthamoeba-keratitis
- Source :
- PLoS ONE, PLoS ONE, Vol 17, Iss 1, p e0262223 (2022), PLoS ONE, Vol 17, Iss 1 (2022)
- Publication Year :
- 2022
- Publisher :
- Public Library of Science, 2022.
-
Abstract
- Contact lens usage has contributed to increased incidence rates of Acanthamoeba keratitis (AK), a serious corneal infection that can lead to blindness. Since symptoms associated with AK closely resemble those incurred by bacterial or fungal keratitis, developing a diagnostic method enabling rapid detection with a high degree of Acanthamoeba-specificity would be beneficial. Here, we produced a polyclonal antibody targeting the carboxylesterase (CE) superfamily protein secreted by the pathogenic Acanthamoeba and evaluated its diagnostic potential. Western blot analysis revealed that the CE antibody specifically interacts with the cell lysates and conditioned media of pathogenic Acanthamoeba, which were not observed from the cell lysates and conditioned media of human corneal epithelial (HCE) cells, Fusarium solani, Staphylococcus aureus, and Pseudomonas aeruginosa. High titers of A. castellanii-specific antibody production were confirmed sera of immunized mice via ELISA, and these antibodies were capable of detecting A. castellanii from the cell lysates and their conditioned media. The specificity of the CE antibody was further confirmed on A. castellanii trophozoites and cysts co-cultured with HCE cells, F. solani, S. aureus, and P. aeruginosa using immunocytochemistry. Additionally, the CE antibody produced in this study successfully interacted with 7 different Acanthamoeba species. Our findings demonstrate that the polyclonal CE antibody specifically detects multiple species belong to the genus Acanthamoeba, thus highlighting its potential as AK diagnostic tool.
- Subjects :
- Male
Physiology
Staphylococcus
Protozoan Proteins
Antibodies, Protozoan
Acanthamoeba
Pathology and Laboratory Medicine
Biochemistry
Carboxylesterase
Mice
Medical Conditions
Antibody Specificity
Immune Physiology
Medicine and Health Sciences
Staphylococcus Aureus
Enzyme-Linked Immunoassays
Cells, Cultured
Protozoans
Multidisciplinary
Immune System Proteins
Epithelium, Corneal
Eukaryota
Pseudomonas Aeruginosa
Bacterial Pathogens
Helminth Infections
Medical Microbiology
Cytochemistry
Medicine
Pathogens
Immunocytochemistry
Research Article
Neglected Tropical Diseases
Contact Lenses
Science
Immunology
Research and Analysis Methods
Microbiology
Antibodies
Cell Line
Echinococcosis
Pseudomonas
parasitic diseases
Parasite Groups
Parasitic Diseases
Animals
Humans
Trophozoites
Immunoassays
Microbial Pathogens
Bacteria
Organisms
Biology and Life Sciences
Proteins
Epithelial Cells
Cell Biology
Tropical Diseases
Parasitic Protozoans
Early Diagnosis
Acanthamoeba Keratitis
Culture Media, Conditioned
Immunologic Techniques
Immunization
Parasitology
Apicomplexa
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 17
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....9988a545b5f1d8cf27e955a51a0c0eb0