title: Seminested polymerase chain reaction (PCR) for detecting Helicobacter pylori DNA in carotid atheromas.
abstract: A polymerase chain reaction (PCR) method for the detection of the glmM gene, selected as Helicobacter pylori target sequence, was improved. While performing pathogenicity island cagA gene detection to discriminate pathogenic strains in atherosclerotic carotid samples, several cagA-positive but glmM-negative samples were found. Polymorphisms present in the region amplified in the nested PCR reaction could explain this result; primers were therefore designed to perform a seminested reaction; this modification optimized sensitivity while maintaining specificity. A real-time PCR for Helicobacter DNA detection was also setup. The combination of all 4 PCR reactions detected 83% of H. pylori DNA-positive samples in atherosclerotic carotid tissue, 64% of which were cagA gene positive.
authors.affiliation: FLENI, Montaneses 2325, Buenos Aires C1428AQK, Cuidad Autonoma de Buenos Aires, Argentina.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Arias
authors.forename: Horacio
authors.initials: H
authors.lastname: Martinetto
authors.forename: Marcelo
authors.initials: M
authors.lastname: Schultz
authors.forename: Sebastian
authors.initials: S
authors.lastname: Ameriso
authors.forename: Santiago
authors.initials: S
authors.lastname: Rivera
authors.forename: Oscar
authors.initials: O
authors.lastname: Lossetti
authors.forename: Gustavo
authors.initials: G
authors.lastname: Sevlever
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics; D050197:Atherosclerosis / Q000175:diagnosis* / Q000382:microbiology; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002339:Carotid Arteries / Q000382:microbiology; D002340:Carotid Artery Diseases / Q000175:diagnosis* / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis*; D005798:Genes, Bacterial; D044404:Genomic Islands / Q000235:genetics; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D010733:Phosphoglucomutase / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C080866:cagA protein, Helicobacter pylori; C097631:phosphoglucosamine mutase; D010733:Phosphoglucomutase