Seminested polymerase chain reaction (PCR) for detecting Helicobacter pylori DNA in carotid atheromas.
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.
Arias
Eugenia
E
FLENI, Montaneses 2325, Buenos Aires C1428AQK, Cuidad Autonoma de Buenos Aires, Argentina.
Martinetto
Horacio
H
Schultz
Marcelo
M
Ameriso
Sebastian
S
Rivera
Santiago
S
Lossetti
Oscar
O
Sevlever
Gustavo
G
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
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C080866:cagA protein, Helicobacter pylori; C097631:phosphoglucosamine mutase; D010733:Phosphoglucomutase