Helicobacter pylori IgA ELISA


The Helicobacter pylori IgA ELISA is intended for the qualitative screening of IgA autoantibodies to H. pylori in serum or plasma. The Eagle Biosciences Helicobacter pylori IgA ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80558 Categories: ,

Helicobacter Pylori IgA ELISA

Helicobacter Pylori IgA ELISA is For Research Use Only

Size: 1×96 wells
Sensitivity: 1.04 µg/ml
Dynamic Range: 1-200 µg/ml
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 5 µL
Alternative Names: H. Pylori IgA

Assay Background

Helicobacter pylori (H. pylori) is a Gram-negative, micro-aerophilic  bacterium restricted to the stomach and the duodenum. It was identified in 1982 by Australian scientists Barry Marshall and Robin Warren, who found that it was present in patients with chronic gastritis and gastric ulcers, conditions that were not previously believed to have a microbial cause. H. pylori has also been associated as an etiologic agent in the development of duodenal ulcers and stomach cancer. More than fifty percent of the world’s population harbor H. pylori in their upper gastrointestinal tract. However, over 80 percent of individuals infected with this bacterium are asymptomatic and it has been postulated that it may play an important role in the natural stomach ecology. H. pylori is a contagious bacterium, although the exact route of transmission is not known. Person-to-person transmission by either the oral-oral or fecal-oral route is most frequent. Consistent with these transmission routes, the bacteria have been isolated from feces, saliva and dental plaques of some infected patients. Transmission occurs mainly within families in developed nations, yet can also be acquired from the community in developing countries.

Assay Principle

The Helicobacter pylori IgA ELISA employs the qualitative enzyme immunoassay technique. Helicobacter antigen has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Ab present is bound by the immobilized antigen. After washing away any unbound substances, a HRP-conjugated anti human antibody is added to each well and incubate. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Ab bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450 nm ±2 nm. The concentration of Ab in the sample is then determined by comparing the O.D of samples to the standard curve.

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Additional Information

Assay Procedure

  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 100 μl of diluted samples, Calibrators and controls into wells. Leave one well empty for the substrate blank.
  3. Incubate for 60 minutes at RT.
  4. Aspirate each well and wash, repeating the process 2 times for a total 3 washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  5. Add 100 μl 1X Antibody solution into each well (expect the substrate blank). Incubate for 30 minutes at RT.
  6. Wash as according to step 4.
  7. Add 100 μl of TMB Reagent to each well. Incubate for 20 minutes at room temperature.
  8. Add 100 μl of Stop Solution to each well.
  9. Read the OD with a microplate reader at 450 nm immediately.

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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