title: A sensitive and specific PCR method to detect Helicobacter felis in a conventional mouse model.
abstract: Although many detection methods have been used to determine Helicobacter colonization in small animal models, the sensitivity and specificity of these detection methods are limited. To improve the Helicobacter felis conventional mouse model for accurate evaluation of therapeutic regimens, we developed a PCR for detection of, and a competitive PCR for quantitation of, H. felis in viral antibody-free (VAF) mice. The PCR was based on the H. felis 16S rRNA gene. An internal control DNA was used for competitive quantitation of the PCR. VAF conventional Swiss-Webster mice were infected with an H. felis culture by oral gavage. At various times after H. felis challenge and therapy, stomach mucosa was collected and evaluated by PCR. PCR detected approximately 50 to 100 H. felis cells per mouse stomach and showed no cross-reaction with other bacteria commonly found in mouse stomachs. Colonization of H. felis in the mouse stomach was confirmed by culture isolation from germfree mice and histological examination of VAF mice. Response to therapy in this H. felis model correlated well with results seen in human clinical trials with H. pylori. A model utilizing PCR detection which may be useful for discovering new antibiotics and/or vaccines against Helicobacter ulcer disease has been developed.
authors.affiliation: Department of Enzymology, Merck Research Laboratories, Merck and Co., Inc, Rahway, New Jersey 07065-0900, USA.
authors.forename: L
authors.initials: L
authors.lastname: Kong
authors.forename: J G
authors.initials: JG
authors.lastname: Smith
authors.forename: D
authors.initials: D
authors.lastname: Bramhill
authors.forename: G K
authors.initials: GK
authors.lastname: Abruzzo
authors.forename: C
authors.initials: C
authors.lastname: Bonfiglio
authors.forename: C
authors.initials: C
authors.lastname: Cioffe
authors.forename: A M
authors.initials: AM
authors.lastname: Flattery
authors.forename: C J
authors.initials: CJ
authors.lastname: Gill
authors.forename: L
authors.initials: L
authors.lastname: Lynch
authors.forename: P M
authors.initials: PM
authors.lastname: Scott
authors.forename: L
authors.initials: L
authors.lastname: Silver
authors.forename: C
authors.initials: C
authors.lastname: Thompson
authors.forename: H
authors.initials: H
authors.lastname: Kropp
authors.forename: K
authors.initials: K
authors.lastname: Bartizal
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics; D004195:Disease Models, Animal; D004359:Drug Therapy, Combination / Q000627:therapeutic use; D005260:Female; D005753:Gastric Mucosa / Q000382:microbiology / Q000473:pathology; D005856:Germ-Free Life; D016998:Helicobacter / Q000235:genetics* / Q000302:isolation & purification*; D016481:Helicobacter Infections / Q000188:drug therapy / Q000382:microbiology / Q000473:pathology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D000900:Anti-Bacterial Agents; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S