Detection of Acanthamoeba spp. using carboxylesterase antibody and its usage for diagnosing Acanthamoeba-keratitis.
Acanthamoeba
/ classification
Acanthamoeba Keratitis
/ diagnosis
Animals
Antibodies, Protozoan
/ analysis
Antibody Specificity
Carboxylesterase
/ administration & dosage
Cell Line
Cells, Cultured
Contact Lenses
/ parasitology
Culture Media, Conditioned
/ metabolism
Early Diagnosis
Epithelial Cells
/ cytology
Epithelium, Corneal
/ cytology
Humans
Immunization
Male
Mice
Protozoan Proteins
/ administration & dosage
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2022
2022
Historique:
received:
30
09
2021
accepted:
20
12
2021
entrez:
5
1
2022
pubmed:
6
1
2022
medline:
19
2
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 34986189
doi: 10.1371/journal.pone.0262223
pii: PONE-D-21-31495
pmc: PMC8730387
doi:
Substances chimiques
Antibodies, Protozoan
0
Culture Media, Conditioned
0
Protozoan Proteins
0
Carboxylesterase
EC 3.1.1.1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0262223Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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