Measurement of Microbial DNA Polymerase Activity Enables Detection and Growth Monitoring of Microbes from Clinical Blood Cultures

Abstract

Surveillance of bloodstream infections (BSI) is a high priority within the hospital setting. Broth-based blood cultures are the current gold standard for detecting BSI, however they can require lengthy incubation periods prior to detection of positive samples. We set out to demonstrate the feasibility of using enzymatic template generation and amplification (ETGA)-mediated measurement of DNA polymerase activity to detect microbes from clinical blood cultures. In addition to routine-collected hospital blood cultures, one parallel aerobic blood culture was collected and immediately refrigerated until being transported for ETGA analysis. After refrigeration holding and transport, parallel-collected cultures were placed into a BACTEC incubator and ETGA time-course analysis was performed. Of the 308 clinical blood cultures received, 22 were BACTEC positive, and thus were initially selected for ETGA time course analysis. The ETGA assay detected microbial growth in all 22 parallel-positive blood cultures in less time than a BACTEC incubator and also yielded genomic DNA for qPCR-based organism identification. In summary, feasibility of detecting microbes from clinical blood culture samples using the ETGA blood culture assay was demonstrated. Additional studies are being considered towards development of clinically beneficial versions of this methodology.

Systemic and Extraradicular Bacterial Translocation in Apical Periodontitis

“…Although bacterial cultures are considered the reference method to determine the presence of microorganisms ( Zweitzig et al., 2013 ), molecular analysis is more sensitive, especially in anaerobic conditions, such as necrotized root canals ( Baumgartner, 2004 ). Particularly, Porphyromonas is considered fastidious genera and culture methods may underestimate their frequency ( Gomes and Herrera, 2018 ).…”

Serum bacterial DNA detection in patients with cholangitis after Kasai procedure

“…14,15 Because only 20% of all bacteria can be cultured using conventional techniques, it is not always easy to make a prompt and accurate diagnosis of cholangitis, although blood cultures can directly detect the organism causing cholangitis. Several studies showed that PCR-based methods were more sensitive than standard cultures for detecting translocated bactDNA in blood samples, [16][17][18] whereas, up to now, studies of bactDNA in patients with cholangitis have rarely been reported. We found that translocation of bactDNA is a frequent event in patients with cholangitis.…”