Abstract
BackgroundSepsis is one of the main causes of mortality and morbidity. The rapid detection of pathogens in blood of septic patients is essential for adequate antimicrobial therapy and better prognosis. This study aimed to accelerate the detection and discrimination of Gram-positive (GP) and Gram-negative (GN) bacteria and Candida species in blood culture samples by molecular methods.MethodsThe Real-GP®, -GN®, and -CAN® real-time PCR kit (M&D, Wonju, Republic of Korea) assays use the TaqMan probes for detecting pan-GP, pan-GN, and pan-Candida species, respectively. The diagnostic performances of the real-time PCR kits were evaluated with 115 clinical isolates, 256 positive and 200 negative blood culture bottle samples, and the data were compared to results obtained from conventional blood culture.ResultsEighty-seven reference strains and 115 clinical isolates were correctly identified with specific probes corresponding to GP-bacteria, GN-bacteria and Candida, respectively. The overall sensitivity and specificity of the real-time PCR kit with blood culture samples were 99.6% and 89.5%, respectively.ConclusionsThe Real-GP®, -GN®, and -CAN® real-time PCR kits could be useful tools for the rapid and accurate screening of bloodstream infections (BSIs).
Highlights
Sepsis is one of the main causes of mortality and morbidity
Sensitivity and specificity of the Real-GP®, -GN®, and -CAN® real-time PCR TaqMan assay with reference bacterial and fungal strains The detection limit of the real-time PCR TaqMan assay for GP, GN-bacteria, and Candida was 103 CFU/mL, CFU/mL, and CFU/mL, respectively
Results of Real-GP®, -GN®, and -CAN® real-time PCR TaqMan assay with clinical isolates The results between subculture and real-time PCR assay were completely concordant (100%) in 115 clinical isolates
Summary
Sepsis is one of the main causes of mortality and morbidity. The rapid detection of pathogens in blood of septic patients is essential for adequate antimicrobial therapy and better prognosis. This study aimed to accelerate the detection and discrimination of Gram-positive (GP) and Gram-negative (GN) bacteria and Candida species in blood culture samples by molecular methods. Gram-positive (GP) bacteria are the most frequent causative agents of bloodstream infections (BSIs) (30-50% of all cases), followed by Gram-negative (GN) bacteria in 25-30% and fungal infections representing 1-3% of all sepsis cases [4]. Some PCR-based assays could identify specific bacterial pathogens [10,11], while broad range bacterial PCR can detect almost any bacterial species [15,16]. Broad-range bacterial PCR has a great advantage in that it is able to detect microorganisms that are found less frequently, or can even identify unknown causative agents of bacterial origin
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