ABSTRACT. Urea breath test (UBT) using an infrared spectral analyzer is widely used for non-invasive and rapid detection of gastric Helicobacter spp. in human, but not veterinary medicine. The main purposes of this study were to determine the reference range of the UBT in dogs and to evaluate its clinical usefulness. To address the first aim, 6 healthy laboratory beagles were subjected to UBT and upper gastrointestinal endoscopy. Gastric endoscopic biopsy samples from the antrum, corpus and fundus were examined for Helicobacter spp. by polymerase chain reaction (PCR) testing, rapid urease test (RUT), histology and cytology. Amoxicillin, metronidazole and omeprazole were given to infected dogs for 14 days, and dogs that became Helicobacter-negative were used to determine the reference range for UBT. To address the second aim, 32 canine patients underwent UBT before upper gastrointestinal endoscopy, and the sensitivity and specificity of UBT were calculated based on our newly determined reference range using PCR as the gold standard for detection of Helicobacter spp. Initially, all 6 laboratory beagles were infected in all gastric regions and became uninfected after eradication. The mean ± 2 SD UBT value after eradication was 0.6 ± 1.8‰, and the reference range for UBT was determined to be less than 2.5‰. UBT was completed successfully in 27 patients. Using our reference range, UBT displayed 89% (16/18) sensitivity and 89% (8/9) specificity, indicating that UBT was quite useful for the detection of gastric Helicobacter spp. infection in dogs. Helicobacter spp. colonizes the stomach and intestine of humans and several animal species [8]. The Helicobacter genus currently includes about 38 formally named members with numerous other putative species under investigation [11]. In humans, Helicobacter pylori is known to be the major agent of chronic diffuse superficial gastritis, plays a causative role in peptic ulcers and is considered a co-factor in the development of gastric malignancies [15,28,36] [4,12], but insufficient epidemiological data are available to estimate the prevalence in Japan. In addition, although H. felis, H. bizzozeronii, H. salomonis and "Candidatus Helicobacter heilmannii" are also found in the human stomach [3,38,40] and are suggested to be associated with gastric diseases in humans [18,19,37,42], their pathogenic significance in dogs is controversial [10,16,[29][30][31][32][33][34].Diagnostic tests for gastric Helicobacter spp. in dogs and cats include polymerase chain reaction (PCR) testing, rapid urease test (RUT), histology and cytology; all of these require anesthesia and gastric biopsy [9,21,22]. The 13 C-urea breath test (UBT) is a non-invasive test with high sensitivity and specificity that is widely used in human medicine [7,14,23,35]. In a multicenter trial in Japan, UBT using an infrared spectral analyzer showed 98.1% sensitivity and 97.9% specificity in human when a cut-off value of 2.5‰ was used to distinguish between patients with and without Helicobacter infections [23]. In veteri...