Absence of Bacteria on Coronary Angioplasty Balloons from Unselected Patients: Results with Use of a High Sensitivity Polymerase Chain Reaction Assay.

Gorm Mørk Hansen,Michael Givskov,Peter Riis Hansen,Claus Henrik Nielsen,Steffen Helqvist,Martin Nilsson,Tim Tolker-Nielsen,Palle Holmstrup,Özlem Yilmaz

doi:10.1371/journal.pone.0145657

Abstract

Periodontitis is a chronic, bacterially-induced inflammatory disease of the tooth-supporting tissues, which may result in transient bacteremia and a systemic inflammatory response. Periodontitis is associated with coronary artery disease independently of established cardiovascular risk factors, and translocation of bacteria from the oral cavity to the coronary arteries may play a role in the development of coronary artery disease. Very few studies have used angioplasty balloons for in vivo sampling from diseased coronary arteries, and with varying results. Therefore, the aim of this study was to assess if bacterial DNA from primarily oral bacteria could be detected on coronary angioplasty balloons by use of an optimized sampling process combined with an internally validated sensitive polymerase chain reaction (PCR) assay. Coronary angioplasty balloons and control samples from a total of 45 unselected patients with stable angina, unstable angina/non-ST elevation myocardial infarction, and ST-elevation myocardial infarction (n = 15 in each group) were collected and analyzed using a PCR assay with high sensitivity and specificity for 16S rRNA genes of the oral microbiome. Despite elimination of extraction and purification steps, and demonstration of sensitivity levels of 25–125 colony forming units (CFU), we did not detect bacterial DNA from any of the coronary angioplasty balloons. A subsequent questionnaire indicated that the prevalence of periodontitis in the study cohort was at least 39.5%. Although coronary angioplasty balloons are unlikely to be useful for detection of bacteria with current PCR techniques in unselected patients with coronary artery disease, more studies are warranted to determine the extent to which bacteria contribute to atherosclerosis and its clinical manifestations and whether the presence of bacteria in the arteries is a transient phenomenon.

Highlights

Cardiovascular disease (CVD) is the leading cause of death worldwide and its primary cause, atherosclerosis, is a chronic inflammatory disease of the arteries, which is strongly associated with risk factors such as hypertension, smoking, diabetes, obesity, hyperlipidemia, a family history of CVD, and socioeconomic deprivation
Detection limits were between 25–125 colony-forming units (CFU) for angioplasty balloons dipped in blood spiked with a Gram negative bacterium (P. aeruginosa), depending on the amount of blood (1–5 μL) adhering to the angioplasty balloon
We developed a highly sensitive polymerase chain reaction (PCR)-based assay that was capable of detecting 25–125 colony forming units (CFU) and theoretically, a similar number of single bacteria adhering to angioplasty balloons with optimized handling of samples by elimination of extraction and purification steps

Summary

Introduction

Cardiovascular disease (CVD) is the leading cause of death worldwide and its primary cause, atherosclerosis, is a chronic inflammatory disease of the arteries, which is strongly associated with risk factors such as hypertension, smoking, diabetes, obesity, hyperlipidemia, a family history of CVD, and socioeconomic deprivation. Several large randomized controlled studies with antibiotic treatment have failed to demonstrate improved cardiovascular outcomes and mortality in patients with coronary artery disease, and more recently it has been suggested that organization of bacteria in antibiotic-resistant biofilms may have contributed to these negative results [29,30,31,32,33,34]

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