The utility of Helicobacter pylori serology was evaluated in 4,722 specimens and compared to stool antigen detection. Immunoglobulin M (IgM) sensitivity (6.8%) was unacceptably low. Key performance differences were observed in IgG specificity, IgA sensitivity, and specificity between adults and children that may warrant differentiating optimal serologic cutoff values by age.Helicobacter pylori causes gastrointestinal disease in both children and adults (10). Noninvasive diagnostic tests include the [13 C]urea breath test, serology, and stool antigen testing (HpSA). Numerous studies have evaluated these diagnostic tests but have been limited by a small sample size or restriction to either children or adults (3-9, 11-17, 19, 20). The clinical utility of serologic testing in both children and adults has been debated; moreover, it has not been established whether positive cutoff levels need to be adjusted for age (4,5,8,13,19). Immunoglobulin A (IgA) and IgG serologic tests are possibly less reliable in children than adults, but this has not been definitively established (13). Some investigators have supported the use of IgM as an indicator of active disease (2), while others have found IgM to have little diagnostic utility (7,18). Because of the conflicting data, we performed a large-scale study on H. pylori serology to analyze its utility and differences in performance in children and adults.Paired results of H. pylori serology (IgG, IgA, and/or IgM) and HpSA from October 1998 to January 2009 were analyzed in tests performed within 2 months of each other. HpSA was performed using the Premier Platinum HpSA Plus enzyme immunoassay according to the manufacturer's instructions (Meridian Bioscience, Inc., Cincinnati, OH). The cutoff optical density at 450 nm was Ͻ0.100 for a negative result and Ն0.100 for a positive result.Serology has been performed with in-house enzyme-linked immunosorbent assay (ELISA) kits used since 1998. The IgG and IgA ELISAs were validated against the Enteric Products Inc. (Stony Brook, NY) ELISA. The IgM ELISA was validated against the MRL (now Focus Diagnostics, Cypress, CA) IgM ELISA. H. pylori antigens (CagA and VacA; Micro Detect, Inc., Tustin, CA) were used to coat microtiter plates at 1.0 g/ml. Samples were diluted 1:101 for IgG and IgA and 1:51 for IgM and then reacted at room temperature for 30 min. After washing, diluted horseradish peroxidase-conjugated antihuman IgG, IgA, or IgM was reacted for 30 min at room temperature. After washing again, the wells were developed with tetramethylbenzidine for 30 min and the absorbance was measured at 450 nm. The cutoffs (in index values) for IgG and IgA were Յ1.7 for a negative result, 1.8 to 2.2 for an equivocal * Corresponding author. Mailing address: 500 Chipeta Way, ARUP Laboratories, Salt Lake City, UT 84108. Phone: (801)