Microtiter Wells Elisa Test Kit , Reliable H.Pylori Test Kit 60 ML Washing Buffer

H.Pylori IgG Antibody ELISA Test kit,96wells/kit, Elisa Sandwich method, for quantitative measurement

Intended Use:

The HAV IgG/IgM Rapid Test is a lateral flow chromatographic immunoassay for the simultaneous detection and differentiation of IgG and IgM antibodies to Hepatitis A virus (HAV) in human serum, plasma or whole blood. It is intended to be used by professionals as a screening test and as an aid in the diagnosis of infection with HAV. Any reactive result with the HAV IgG/IgM Rapid Test must be confirmed with alternative testing method(s) and clinical findings.

Summary:

H. pylori is a small, spiral-shaped bacterium that lives in the surface of the stomach and duodenum. It is associated in the etiology of a variety of gastrointestinal diseases including duodenal and gastric ulcer, non-ulcer dyspepsia and active and chronic gastritis.1,2 H. pylori infection is present in over 90% of duodenal ulcers, 80% of gastric ulcers, and 70% of gastritis. Recently, it has been classified as a Class I carcinogen by the WHO. Although the transmission route for H. pylori is not yet known, it is believed to be transmitted by oral-oral or fecal-oral route. Both invasive and non-invasive methods are used to diagnose H. pylori infection in patients with symptoms of gastrointestinal disease. Specimen-dependent and costly invasive diagnostic methods include gastric or duodenal biopsy followed by urease testing (presumptive), culture, and/or histologic staining.3 Non-invasive techniques include the urea breath test, which requires expensive laboratory equipment and moderate radiation exposure, and serological methods.4,5 Individuals infected with H. pylori develop serum antibodies which correlate strongly with histologically confirmed H. pylori infection.6,7,8 Early in the course of active infection, IgM antibody levels may be detectable. Levels of IgG and IgA antibody rise with the infection and remain constantly high until infection is eliminated. Therefore, the efficacy of antimicrobial therapy can be monitored by detecting changes in the antibody levels. The H. pylori IgG EIA Test Kit is an immunoassay for the qualitative and quantitative detection of the presence of IgG antibodies to H. pylori in serum or plasma specimen. The test utilizes recombinant H. pylori antigens to selectively detect IgG antibodies to H. pylori in serum or plasma.

Regents Provided

TEST PRINCIPLE

The H. pylori IgG EIA Test Kit is a solid phase enzyme immunoassay based on indirect principle for the qualitative and quantitative detection of IgG antibodies to H. pylori in human serum or plasma. The microwell plate is coated with H. pylori recombinant antigens. During testing, the specimen diluent and the specimens are added to the antigen coated microwell plate and then incubated. If the specimens contain IgG antibodies to H. pylori, it will bind to the antigens coated on the microwell plate to form immobilized antigen-H. pylori IgG antibody complexes. If the specimens do not contain IgG antibodies to H. pylori, the complexes will not be formed. After initial incubation, the microwell plate is washed to remove unbound materials. The enzyme-conjugated anti-human IgG antibodies are added to the microwell plate and then incubated. The enzyme-conjugated anti-human IgG antibodies will bind to the immobilized antigen-H. pylori IgG antibody complexes present. After the second incubation, the microwell plate is washed to remove unbound materials. Substrate A and substrate B are added and then incubated to produce a blue color indicating the amount of H. pylori IgG antibodies present in the specimens. Sulfuric acid solution is added to the microwell plate to stop the reaction producing a color change from blue to yellow.

TEST PROCEDURE

1.  Prepare a sufficient amount of microtiter wells for the standards, controls and samples as well as for a substrate blank.
2.  Pipet 100 µL each of the diluted (1:101) samples and the ready-to-use standards and controls respectively into the wells. Leave one well empty for the substrate blank.
3.  Cover plate with the re-usable plate cover and incubate at room temperature for 60 minutes.
4.  Empty the wells of the plate (dump or aspirate) and add 300 µL of diluted washing solution. This procedure is repeated totally three times. Rests of the washing buffer are afterwards removed by gentle tapping of the microtiter plate on a tissue cloth.
5.  Pipet 100 µL each of ready-to-use conjugate into the wells. Leave one well empty for the substrate blank.
6.  Cover plate with the re-usable plate cover and incubate at room temperature for 30 minutes.
7.  Empty the wells of the plate (dump or aspirate) and add 300 µL of diluted washing solution. This procedure is repeated totally three times. Rests of the washing buffer are afterwards removed by gentle tapping of the microtiter plate on a tissue cloth.
8.  Pipet 100 µL each of the ready-to-use substrate into the wells. This time also the substrate blank is pipetted.
9.  Cover plate with the re-usable plate cover and incubate at room temperature for 20 minutes in the dark (e.g. drawer).
10.  To terminate the substrate reaction, pipet 100 µL each of the ready-to-use stop solution into the wells. Pipet also the substrate blank.
11.  After thorough mixing and wiping the bottom of the plate, perform the reading of the absorption at 450 nm (optionally reference wavelength of 620 nm). The color is stable for at least 60 minutes.

ASSAY CHARACTERISTICS