[Cloning, expression and immunodiagnostic evaluation of antigen EPC1 from Echinococcus granulosus].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
; 29(3): 167-71, 2011 Jun.
Article
em Zh
| MEDLINE
| ID: mdl-21970102
ABSTRACT
OBJECTIVE:
To clone and express EPCl gene of Echinococcus granulosus, and investigate its immunogenicity and diagnostic value.METHODS:
Total RNA was extracted from hydatid cyst protoscoleces and EPC1 gene of Echinococcus granulosus was amplified by RT-PCR. The PCR product was cloned into pGEM-T vector, and then subcloned into the prokaryotic expression vector PET28a(+). The positive recombinants were transformed into Escherichia coli BL21 (DE3), and followed by expression of the protein induced by IPTG. The recombinant protein was identified by SDS-PAGE and Western blotting, and used to establish ELISA. Serum samples from patients with cystic echinococcosis (60 cases), alve-olar echinococcosis (37 cases), cysticercosis (16 cases), clonorchiasis sinensis (7 cases), schistosomiasis japonica (4 cases) and healthy persons (33 cases) were examined.RESULTS:
The recombinant plasmid PET28a-EgEPC1 was identified by restriction enzyme digestion and sequencing. SDS-PAGE result showed that the recombinant containing recombinant plasmid PET28a-EgEPC1 expressed a soluble fission protein of EgEPC1 (about M, 11 000). The protein was recognized by pool sera of cystic echinococcosis patients. The overall sensitivity and specificity of diagnosis by ELISA for cystic echinococcosis were 78.3% (47/60), and 98.3% (59/60), respectively. The cross reaction with sera of alveolar echinococcosis was 40.5% (15/37).CONCLUSION:
The recombinant EgEPC1 antigen has diagnostic value in cystic echinococcosis.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Helminto
/
Echinococcus granulosus
/
Equinococose
/
Antígenos de Helmintos
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
Assunto da revista:
PARASITOLOGIA
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
China