Previously, immunoglobulin G (IgG) antibodies to five antigens with a relative molecular mass of between 15 and 30 kDa from Helicobacter pylori were found to be significantly more frequent in H. pylori-infected patients than in noninfected patients. In this study, these specific low-molecular-mass (LMW) antigens were separated by ultrafiltration of whole-cell sonicates. The LMW antigen preparation was evaluated by enzymelinked immunosorbent assay with serum samples from 76 children with abdominal symptoms and 151 adults with dyspeptic symptoms. H. pylori was cultured or seen in 40 (53%) children and 83 (55%) adults. Increased antibody levels to H. pylori were found in serum from 35 (46%) children and 88 (58%) adults. Values for sensitivity, specificity, and predictive value of positive and negative results of the test were higher with LMW antigens than with the heat-stable antigen previously described. The low specificity and predictive value of a positive result were due to seropositive results for 21 persons with a negative culture for H. pylori and negative microscopy results for Helicobacter-like organisms in biopsies from gastric mucosa. Histologically, chronic gastritis was demonstrated in 43% of these persons, and 19% had peptic ulcer, indicating that they have or have had H. pylori infection. Specific antibodies to H. pylori were confirmed in all 21 patients by the Western immunoblot technique. Use of the LMW antigen improved the IgG antibody detection in patients with H. pylori infection, even though the results reflect the difficulties in establishing a true gold standard for diagnosis of H. pylori infection.
Helicobacter pylori from patients with different diseases, including so-called autoimmune thyroiditis, chronic tonsillitis and tonsillar cancer, was isolated and cultured. It was identified according to the genotype using labeled hybridization probes complementary to six sequences of cagA and vacA genes. Different types of strains were found in isolates from gastrointestinal tract and patients suffering from thyroiditis. Six out of seven genotyped isolates from patients in our Department of Otorhinolaryngology and Head and Neck Surgery exhibited the same genotype, differing from isolates obtained from other patients; the 7th isolate originated from a patient who had undergone surgery for deviatio septi nasi, at the same time suffering from autoimmune thyroiditis, having confirmed gastric infection by H. pylori from biopsy. This data made it possible to formulate the hypothesis on probable association of specific H. pylori genotype with chronic tonsillitis and tonsillar cancer. We assessed commercial transport media and improved nucleic acid isolation techniques and the RT-PCR-based tests, which allowed us to skip a culture step and to test directly the patients' samples; however, for full confirmation of our hypothesis and explanation of possible mechanisms of the contribution of Helicobacter sp. to the pathogenesis of the disease further data are to be collected and evaluated.
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