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.

Extragastric Manifestations of Helicobacter pylori Infection – Other Helicobacters

“…Arias et al. utilizing a seminested polymerase chain reaction (PCR), detected H. pylori DNA in 83% of the atherosclerotic carotid tissue samples, 64% of which were cagA positive [10], while Weiss et al. did not find evidence for a direct role of H. pylori nor Mycoplasma pneumoniae in carotid artery atherosclerosis [11].…”

A Systematic Review and Meta-Analysis on the Relation Between Helicobacter Pylori Infection and Atherosclerosis in the Iranian Population

“…For instance, the rate of isolation of H. pylori from atherosclerosis was 27.2-33.5% in the study by Jha et al [92]. In another study in Argentina, the carotid plaque isolation was about 83% [95]. In a cross-sectional study in Turkey, Kilic et al found that the rates of H. pylori isolation from the atherosclerotic plaques and non-atherosclerotic vascular wall specimens were 48.2% and 19.2%, respectively [96].…”

Association between Helicobacter pylori infection and carotid atherosclerosis in Chinese adults

“…H. pylori may contribute to atherosclerosis disease through several pathways. It might directly invade the vessel wall and induce dyslipidemia [ 5 , 6 , 35 ]. Chronic H. pylori infection could trigger an immune response.…”

“…Stroke is the second leading cause of death and a major cause of disability worldwide [ 1 ]; carotid atherosclerosis plays a fundamental role in the occurrence of ischemic stroke [ 2 ]; atherosclerosis is considered a chronic inflammation-driven disease of the arterial wall [ 3 ]. Persistent infectious pathogens, such as Helicobacter pylori ( H. pylori ), have the possibility of promoting the development of atherosclerosis [ 4 ] because H. pylori can either directly invade vessel walls [ 5 ] or indirectly induce endothelial dysfunction and dyslipidemia [ 6 ]. Thus, the association between H. pylori infection and atherosclerosis disease has received considerable attention [ 7 , 8 ].…”