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Anfahrt




Product Name: Antibody to Hepatitis E Virus (Total) ELISA Kit 

Product Code: HEV-HB001

Principle: Two-step Incubation, Double Antigen Sandwich

Intended Use

This HEV-Ab ELISA kit is an enzyme linked-immunosorbent assay for in vitro qualitative detection of total antibodies (IgG, IgM,etc.) to hepatitis E virus in animal serum or plasma. It is intended for use in medical laboratories for the research of infection among the population and zoonosis related research on hepatitis E virus.

Principle of the assay

This HEV-Ab ELISA kit uses polystyrene microwell strips pre-coated with recombinant HEV antigens (HEV-Ag) corresponding to structural proteins ORF-2 of the native virus. Serum or plasma sample is added into the microwells. In case of presence of HEV-Ab in the sample, the pre-coated antigens will be bound to the antibody and during the first incubation step, the specific immunocomplex formed is captured on the solid phase. After washing to remove unbound sample, second recombinant HEV antigen conjugated to Horseradish Peroxidase (HRP) is added into the wells. During the second incubation step, this antigen will bind to the second variable domain of the HEV antibodies if they have been captured by HEV-antigen during the first incubation step. The unbound HRP conjugate is removed during washing and Chromogen solutions containing Tetramethylbenzidine (TMB) and urea peroxide are added into the wells. In presence of the antigen-antibody-antigen (HRP) "sandwich" complex, the colorless Chromogens are hydrolyzed by the bound HRP-Conjugate to a blue colored product. The amount of color intensity can be measured and is proportional to the amount of antibody captured in the wells, and to the sample respectively. Wells containing samples negative for HEV remain colorless.

Assay procedure:

  • Add sample diluent            100µl
  • Add sample                         10µl
  • Incubate                              30minutes
  • Wash                                   5 times
  • Add HRP-Conjugate            100µl
  • Incubate                              30minutes
  • Wash                                   5 times
  • Coloring                              50µl A + 50µl B
  • Incubate                             10minutes
  • Stop the reaction                50µl stop solution
  • Read the absorbance 450nm or 450/630 nm

Major Components:

  • Microwell plate                      One/96 wells
  • Negative/Positive control        One each/ 0.5ml
  • Sample diluent                      One/ 6.0ml
  • HRP-Conjugate                     One/12ml
  • Wash Buffer                          One/50ml
  • Chromogen A/B Stop solution One each/7ml