Related Experiment Videos
Hepatitis-E-virus-associated antigen: improved detection in stools by protein Fv removal
A Diallo1, Y Lazizi, B Le Guenno
1Laboratoire de Virologie, Institut Pasteur, Dakar.
Research in Virology
|November 1, 1991
Summary
Detecting hepatitis E virus antigen (HEV-AAg) in stools was improved by minimizing interference from protein Fv. This enhanced assay accurately detects HEV-AAg and can also identify serum antibodies to HEV.
Area of Science:
- Hepatology
- Virology
- Immunology
Background:
- Hepatitis E virus-associated antigen (HEV-AAg) detection in stools can be affected by protein Fv, a factor released during infectious hepatitis.
- Simultaneous appearance of transaminase elevation, HEV-AAg shedding, and Fv leakage was observed in an infected monkey model.
Purpose of the Study:
- To reappraise ELISA detection of HEV-AAg in stools, addressing interference from protein Fv.
- To develop an improved, specific assay for HEV-AAg detection and explore its utility for detecting HEV antibodies.
Main Methods:
- Comparison of labeled human IgG and pre/post-inoculation simian IgG for HEV-AAg and Fv detection.
- Evaluation of coated human IgM and pre/post-inoculation simian IgM in capture assays.
- Utilizing simian IgM and beta-galactosidase-labeled simian IgG to minimize Fv interference.
- Adsorption with monoclonal IgM to ensure HEV specificity.
- Testing the improved assay on stool samples from Senegalese patients with viral hepatitis.
Main Results:
- Simian IgM and beta-galactosidase-labeled simian IgG reduced Fv interference, showing promise for HEV-AAg detection.
- Despite improvements, Fv still caused false positives, necessitating monoclonal IgM adsorption for specificity.
- HEV-AAg was detected in 6 out of 30 stool samples from patients with viral hepatitis.
- No HEV-AAg positivity was found in patients with hepatitis A, B, cytomegalovirus, or Epstein-Barr virus.
- Inhibition experiments confirmed assay specificity and demonstrated its utility for detecting serum antibodies to HEV-AAg.
Conclusions:
- An improved ELISA method using simian IgM and monoclonal IgM adsorption enhances HEV-AAg detection specificity in stools.
- The developed assay successfully identified HEV-AAg in a subset of viral hepatitis patients and differentiated it from other hepatitis types.
- This assay is valuable for diagnosing HEV infections and can be adapted to detect serum antibodies against HEV-AAg.