ELISA technique standardization for strongyloidiasis diagnosis

Objective: To standardize ELISA technique for human Strongyloides stercoralis infection diagnosis. Material and methods: A crude antigen was prepared using filariform larvae obtained from positive stool samples cultured with charcoal. Harvested larvae were crushed by sonication and washed by centrif...

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Detalles Bibliográficos
Autores: Huapaya, Pedro, Espinoza, Yrma, Huiza, Alina, Sevilla, Carlos
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2002
País:Perú
Institución:Universidad Nacional Mayor de San Marcos
Repositorio:Revistas - Universidad Nacional Mayor de San Marcos
Idioma:español
OAI Identifier:oai:revistasinvestigacion.unmsm.edu.pe:article/1496
Acceso en línea:https://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/1496
Access Level:acceso abierto
Palabra clave:Strongyloides stercoralis
estrongiloidiosis
ELISA
serología
diagnóstico de laboratorio
strongyloidiasis
serology
diagnosis laboratory.
Descripción
Sumario:Objective: To standardize ELISA technique for human Strongyloides stercoralis infection diagnosis. Material and methods: A crude antigen was prepared using filariform larvae obtained from positive stool samples cultured with charcoal. Harvested larvae were crushed by sonication and washed by centrifugation in order to obtain protein extracts to be used as antigen. Final protein concentration was 600 µg/mL. Several kinds of ELISA plates were tested and antigen concentration, sera dilution, conjugate dilution and cut off were determined to identify infection. Sera from patients with both hyperinfection syndrome and intestinal infection demonstrated by parasitological examination were positive controls and sera from people living in non-endemic areas with no infection demonstrated by parasitological examination were negative controls. Results: Best values were 5 µg/mL for antigen, 1/64 for sera, 1/1000 for conjugate; optical density values for positive samples were 1,2746 (1,1065 – 1,4206, DS = 0,3284) and for negative samples 0,4457 (0,3324 – 0,5538, DS = 0,2230). Twenty sera samples from positive subjects and one hundred from negative subjects were examined, obtaining 90% sensitivity and 88% specificity. Conclusion: The results show this technique could be useful as strongyloidiasis screening test in population studies.