Production of recombinant 14-3-3 protein and determination of its immunogenicity for application in serodiagnosis of strongyloidiasis.
Trans R Soc Trop Med Hyg
; 113(6): 326-331, 2019 06 01.
Article
en En
| MEDLINE
| ID: mdl-30856271
ABSTRACT
BACKGROUND:
Strongyloides stercoralis is the fourth most important intestinal nematode worldwide. The parasite load and larvae count are often low, thus conventional methods are not sufficiently sensitive to detect the infection. In this study we developed an immunoglobulin G-based enzyme-linked immunosorbent assay (ELISA) method to detect antibodies against S. stercoralis 14-3-3 protein in patients' sera.METHODS:
S. stercoralis RNA was extracted and following complementary DNA synthesis, the 708-bp fragment of 14-3-3 protein was amplified by polymerase chain reaction and cloned into the pET28a+ expression vector. The 30-kDa recombinant 14-3-3 protein was expressed in Escherichia coli BL21 (DE3) cells and purified by affinity chromatography. Finally, its immunoreactivity was assessed by indirect ELISA and western blotting.RESULTS:
The S. stercoralis 14-3-3 gene was successfully amplified and cloned into an expression vector. The 30-kDa recombinant protein was purified by affinity chromatography. An ELISA developed in-house detected infected patients' sera with 96% sensitivity.CONCLUSIONS:
We concluded that the recombinant 14-3-3 protein has enough sensitivity and specificity for detection of strongyloidiasis in human sera and could be applied for serodiagnosis.Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Contexto en salud:
3_ND
/
4_TD
Problema de salud:
3_helminthiasis
/
3_neglected_diseases
/
4_strongyloidiasis
Asunto principal:
Estrongiloidiasis
/
Ensayo de Inmunoadsorción Enzimática
/
Pruebas Serológicas
/
Proteínas 14-3-3
Tipo de estudio:
Diagnostic_studies
/
Observational_studies
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Trans R Soc Trop Med Hyg
Año:
2019
Tipo del documento:
Article
País de afiliación:
Irán