Comparison of the Utility of Recombinant B8/2 Subunit of the Antigen B, Native Antigen, and a Commercial ELISA Kit in the Diagnosis of Human Cystic Echinococcosis

Iran Biomed J. 2019 Jul;23(4):246-52. doi: 10.29252/.23.4.246. Epub 2018 Oct 28.

Abstract

Background: Cystic echinococcosis (CE) is a helminthic disease caused by the larval form of Echinococcus granulosus. In the present study, the B8/2 subunit of antigen B (AgB) of E. granulosus was expressed in E. coli host and then applied in a diagnostic ELISA set up.

Methods: The DNA sequence of AgB8/2 subunit from E. granulosus was extracted from the GenBank and codon-optimized according to E. coli codon usage. The target sequence was cloned in an expression vector (pGEX-4T-1). The produced antigen was used in an ELISA system, and its performance for the diagnosis of human hydatid cyst was evaluated, using sera from CE and non-CE patients, along with the sera from healthy subjects. Moreover, the diagnostic value of the recombinant protein was compared with native AgB, as well as with a commercial kit.

Results: Antibodies to hydatid cyst were detected in 27 out of 30 patients corresponding to a sensitivity of 90% (95% CI: 73-98%). Cross-reaction with sera of non-CE subjects was seen in two cases resulted in a specificity of 93.5% (95% CI: 82-98%) for the test. A sensitivity of 87% and specificity of 90% were found for the native form of the antigen, while the ELISA commercial kit had a sensitivity of 97% and specificity of 95%.

Conclusion: Our data show that rEgAgB8/2 is an appropriate source of antigen for the serological diagnosis of human hydatid cyst. Co-expression of the rEgAgB/2 along with other subunits of AgB may enhance the performances of these antigens for the serodiagnosis of human CE.

Keywords: Diagnosis; Echinococcus granulosus; Hydatid cyst.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Antigens, Helminth / biosynthesis
  • Antigens, Helminth / immunology*
  • Echinococcosis / diagnosis*
  • Echinococcosis / immunology
  • Echinococcosis / parasitology*
  • Echinococcus / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Protein Subunits / biosynthesis
  • Protein Subunits / isolation & purification
  • Protein Subunits / metabolism*
  • Reagent Kits, Diagnostic*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*

Substances

  • Antigens, Helminth
  • Protein Subunits
  • Reagent Kits, Diagnostic
  • Recombinant Proteins