Abstract
Background: This study aimed to evaluate real-time polymerase chain reaction coupled with multiplex probe melting curve analysis (PCR-MCA) for pathogen detection in patients with suspected bloodstream infections (BSIs).Methods: A PCR-MCA assay was developed for simultaneous identification of 28 kinds of the most common pathogens and two resistance genes within a few hours. The diagnostic performance of the PCR-MCA assay was determined and compared to the results of blood culture.Results: A total of 2,844 consecutive new episodes of suspected BSIs in 2,763 patients were included in this study. There were 269 episodes of pathogens identified by blood culture. For all the pathogens tested, the PCR-MCA assay exhibited a sensitivity of 88.8% (239/269), specificity of 100% (2,575/2,575), and agreement of 98.9% (2,814/2,844). For the pathogens on the PCR-MCA list, the PCR-MCA results had a sensitivity of 99.2% (239/241), specificity of 100% (2,575/2,575), and agreement of 99.9% (2,814/2,816) compared with the results of blood culture. For seven samples with multiple pathogens identified simultaneously during one blood culture investigation, the PCR-MCA assay verified the results of the blood culture, with an agreement rate of 100% for each.Conclusion: The PCR-MCA assay could discover 88.8% of the pathogens in clinical practice, showing excellent diagnostic performance vs. that of blood culture for pathogen detection in patients with suspected BSIs, and would contribute to rapid diagnosis and correct antibiotic administration.
Highlights
Sepsis could be called a severe complication of bloodstream infections (BSIs)
The observation unit was an episode of suspected BSI, the results were evaluated as follows: once a bacterial or fungal pathogen was isolated from at least one blood culture, a positive BSI episode was recorded, and the consecutive duplicate results were merged; a BSI episode was considered new if it was detected at least 14 days from the previous infection with positive blood culture or if a different microorganism was isolated from a new blood sample (Quiles et al, 2019); consecutive repeated negative blood culture results from one patient during one hospitalization was recorded as a negative BSI episode
Among the 269 episodes of pathogens identified by blood culture, 241 were identified with a pathogen on the polymerase chain reaction (PCR)-melting curve analysis (MCA) list, while 28 were not
Summary
Sepsis could be called a severe complication of bloodstream infections (BSIs). The reported incidence of sepsis is increasing, and conservative estimates indicate that sepsis is a leading cause of mortality and critical illness worldwide (Singer et al, 2016). Pathogen Detection in Suspected BSIs guidelines (Singer et al, 2016), empirical antibiotic treatment should be given within 1 h once sepsis is suspected, and broadspectrum antibiotics are often recommended. Blood cultures, aiming to detect viable microorganisms in blood, are still considered as the “gold standard” for the microbiological diagnosis of BSIs. Suspected BSIs can be identified if the culture is obtained first and the antibiotic is concomitantly administered within 72 h, whereas if the antibiotic is first and the culture is acquired within 24 h (Singer et al, 2016). This study aimed to evaluate real-time polymerase chain reaction coupled with multiplex probe melting curve analysis (PCR-MCA) for pathogen detection in patients with suspected bloodstream infections (BSIs)
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