Polyacrylamide Gel Electrophoresis of Proteins Method versus Enzyme-Linked Immunosorbent Assay and Immunochromatography with Monoclonal Antibodies in Pediatric Infection with Helicobacter Pylori

The current methods for the diagnosis of Helicobacter pylori infection in children include invasive (direct) methods, that enable the detection of the bacteria or the bacterial urease in gastric biopsies, or noninvasive (indirect) methods, which consist of researching specific antibodies, antigens and urease in other different samples (serum, saliva, urine, stool, exhaled air). The urease functions in H. pylori infection is to neutralize gastric acid by producing NH3 through the hydrolysis of urea. Monochloramine, a NH3-derived compounds has cytotoxic effects on host cells. The Western Blot method refers to the extraction of a a sodium dodecyl sulphate from a Helicobacter pylori strain followed by a separation of the solubilised protein using discontinous polyacrylamide gel electrophoresis according to molecular mass and transfer of the separated proteins to nitrocellulose. The ELISA (enzyme-linked immunosorbent assay) technique refers to the detection of Anti-Helicobacter pylori antibodies type Ig G. We also used an immunochromatographic system with the lateral leakage test strip based on the principle of the sandwich technique with monoclonal antibodies. As a result, more than two types of reactions can be detected on a single assay device by the combination of colloidal gold �labelled antibodies and complementary oligonucleotide-labelled immobilized at different places on a nitrocellulose membrane. Our study aims to assess the performances of the Western Blot method for determining anti-Helicobacter pylori Ig A and Ig G antibodies in correlation with clinical data and other available diagnosis methods. For validation purposes, the results were compared to those obtained using the enzyme-linked immunosorbent assay technique, gastric biopsy and immunochromatography.