Background: Reliable non-invasive methods for detection of Helicobacter pylori infection are required to investigate the incidence, transmission, and clearance of infection in childhood. Aim: To evaluate a new monoclonal enzyme immunoassay (EIA) (FemtoLab H pylori Cnx) for detection of H pylori antigen in stool in a large cohort of children compared with invasive diagnostic methods and the 13 C urea breath test. Patients and methods: A total of 302 symptomatic previously untreated children (aged 0.5-18.7 years; 148 girls) were recruited at three centres. H pylori status was defined by results of culture, histology, the rapid urease test, and the 13 C urea breath test. Stool samples were investigated locally by the EIA using two different production lots. According to the manufacturer's recommendations, an optical density (OD) of 0.150 was used as a cut off value. Results: OD values clearly differentiated between the 92 H pylori infected and the 210 non-infected children (median (5th-95th percentiles) 2.729 (0.232->4.000) v 0.021 (0.009-0.075)). Only two false positive and two false negative results occurred, giving a sensitivity, specificity, positive predictive value, and negative predictive value of 98%, 99%, 98%, and 99%, respectively. No significant relation was found between age and OD values in infected or non-infected children. Conclusions: The monoclonal stool antigen EIA was excellent in diagnosing H pylori infection in symptomatic children. Accuracy was independent of the laboratory, production lot used, or the child's age. Because only 18/116 children <6 years of age were infected with H pylori, further validation of the test is needed in young infected children.

The risk of reinfection with H. pylori is low in children living in Germany. There is no evidence that the reinfection rate depends on the age, sex, or nationality of the child. The low reinfection rate indicates that it is unnecessary to screen or treat asymptomatic family members in order to prevent reinfection.

Reliable non-invasive tests that are feasible in early childhood are essential to the study of transmission of Helicobacter pylori, since most individuals get infected during the first years of life. New tests are validated by comparison with a "gold standard", but no single test for detection of H. pylori infection can be used as a fully reliable reference method. Therefore, concordant results of at least two biopsy-based tests (histology, culture, rapid urease test) are considered as the "gold standard". Most of the validation studies in children included only a few infants and toddlers, with low numbers particularly for H. pylori-infected individuals. Only when increasing numbers of patients were tested and separated into subgroups by age it became apparent that the accuracy of most tests is lower in young children if the same cut-off values are used as established for older children or adults. Therefore, statements such as "a test has been validated with good results in children" must be interpreted with caution, unless different age groups are considered with sufficient numbers of infected and non-infected children in each age group.

SUMMARYAim: To compare the results of culture and epsilometer test with fluorescence in situ hybridization for the detection of Helicobacter pylori and the presence of clarithromycin-susceptible and clarithromycin-resistant strains in antral biopsies from children. Methods: Antral biopsies from 149 unselected children were investigated prospectively; 15 had previously received anti-H. pylori therapy. H. pylori status was defined by histology, rapid urease test and 13 C-urea breath test. Fluorescence in situ hybridization was applied on fresh tissue with probes specific for the clarithromycin-susceptible wild type and three clarithromycin-resistant mutants. Susceptibility to clarithromycin was tested by epsilometer test in two laboratories.Results: Culture and fluorescence in situ hybridization gave negative results in all 66 H. pylori-negative children (specificity, 100%). Of 83 infected children, cultures were successful in 75 (90%), epsilometer test in 71 (86%) and fluorescence in situ hybridization in 77 (93%). Eleven children (13%) showed discrepant results between the applied methods, indicating mixed infection. Clarithromycin-resistant isolates were identified in 16 of 73 previously untreated children. Conclusions: Primary resistance to clarithromycin is common (22%) in H. pylori isolates from children living in Germany. Fluorescence in situ hybridization is an excellent, fast method for the detection of H. pylori and clarithromycin-resistant mutants in gastric biopsies. Multiple biopsies identify mixed infections, indicating that clarithromycin-resistant and clarithromycin-susceptible strains are not evenly distributed within the stomach.