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Infection withHelicobacter hepaticus causes chronic active hepatitis in certain strains of mice and is associated with hepatocellular carcinoma in A/JCr mice. Like the gastric helicobacters,H. pylori andH. mustelae ,H. hepaticus possesses a high level of urease activity. However, theH. hepaticus urease structural gene sequences have not been previously determined, and the role of the urease enzyme in colonization and in pathogenesis is not known. PCR was used to amplify a portion of the urease structural genes fromH. hepaticus genomic DNA. Amplified DNA fragments were cloned, and the nucleotide sequence was determined. The deduced amino acid sequence of the partialH. hepaticus ureA gene product was found to exhibit 60% identity and 75% similarity to the predictedH. pylori UreA. The deduced amino acid sequence of a partialH. hepaticus ureB gene product exhibited 75% identity and 87% similarity to the predictedH. pylori UreB. Diversity amongH. hepaticus isolates was evaluated by means of a restriction fragment length polymorphism (RFLP) assay. The 1.6-kb fragments within theureAB open reading frames, amplified from 11 independent isolates, were digested with the restriction endonucleaseHha I. Three distinct RFLP patterns were observed. Identical RFLP profiles were noted in sequential isolates of one strain ofH. hepaticus during an 18 month in vivo colonization study, suggesting that the urease genes ofH. hepaticus are stable. The urease genes amongH. hepaticus strains were also well conserved, showing 98.8 to 99% nucleotide sequence identity among three isolates analyzed. These findings indicate thatH. hepaticus has urease structural genes which are homologous to those of the gastricHelicobacter species and that these gene sequences can be used in a PCR and RFLP assay for diagnosis of this important murine pathogen.

journal of clinical microbiology

Development of a PCR-Restriction Fragment Length Polymorphism Assay Using the Nucleotide Sequence of the <i>Helicobacter hepaticus</i> Urease Structural Genes <i>ureAB</i>

Development of a PCR-Restriction Fragment Length Polymorphism Assay Using the Nucleotide Sequence of the Helicobacter hepaticus Urease Structural Genes ureAB