An immunofluorescence test for diagnosis of ophthalmic herpes in a mouse corneal model

Abstract

SUMMARYHerpes simplex virus type 1 (HSV-1) ophthalmic disease is the most common cause of corneal blindness in humans worldwide. Current culture techniques for HSV take several days and commercially available HSV laboratory based diagnostic techniques vary in sensitivity. Our study was conducted to evaluate the use of a quicker and simpler method to herpes ophthalmic diagnosis. Corneal smears were made by firm imprints of infected mouse eyes to glass slides, after smears were fixated with cold acetone, and an indirect immunofluorescence (IIF) method was performed using monoclonal antibodies in a murine model of ophthalmic herpes. Eye swabs from infected mice were inoculated in Vero cells for virus isolation. Cytology and histology of the eye were also performed, using hematoxylin-eosin routine. Mouse eyes were examined by slit-lamp biomicroscopy for evidence of herpetic disease at various times postinoculation. We made a comparative evaluation of sensitivity, specificity and speed of methods for laboratory detection of HSV. Our results indicate that this IIF method is quick, sensitive, specific and can be useful in the diagnosis of ophthalmic herpes as demonstrated in an animal model.

Lacaziosis: immunohistochemical evaluation of elements of the humoral response in cutaneous lesions

“…The observation in the present study of the predominance of cells expressing IgG might indicate that lacaziosis, as a chronic infectious disease, suffer a constant antigen stimulus, as previously considered. In frozen histological sections analyzed by immunofluorescence staining, deposits of C3, IgG and IgM were observed in the fungal wall of 100%, 91% and 18% of the lesions, respectively 20 .…”

“…Moreover, fungi with morphological features of unviability were observed in the inflammatory infiltrate. It was previously described that lacaziosis is a disease of long evolution and lesions frequently present many fungal forms that are no longer viable 19 , 20 . We did not perform viability tests in this study, however, as these authors 19 , 20 , we considered the unviability of the fungi in the studied lacaziosis lesions based on their morphological features.…”