scholarly journals Microbiological diagnosis of sepsis: comparison between real-time polymerase chain reaction and blood culture techniques

Critical Care ◽  
2007 ◽  
Vol 11 (Suppl 4) ◽  
pp. P41
Author(s):  
Simona Barnini ◽  
Carlotta Dodi ◽  
Mario Campa
Keyword(s):  
Polymerase Chain Reaction ◽  
Blood Culture ◽  
Real Time ◽  
Microbiological Diagnosis ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Polymerase Chain

Detection of the Escherichia coli pathogenic geneeaewith three real-time polymerase chain reaction methods

2007 ◽  
Vol 53 (3) ◽  
pp. 398-403 ◽  
Author(s):  
Joanne  Karen McCrea ◽  
Chenyi Liu ◽  
Lai-King Ng ◽  
Gehua Wang
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Detection Limit ◽  
Real Time ◽  
Turnaround Time ◽  
Sybr Green ◽  
Sybr Green I ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Polymerase Chain

Several real-time polymerase chain reaction (PCR) methods are currently available to rapidly detect the presence of a specific DNA sequence. When used for detection of pathogenic organisms, the turnaround time for PCR-based methods is much lower than for traditional culture techniques. This study compared the sensitivity of three real-time PCR methods when detecting the Escherichia coli pathogenic gene eae to determine which method is most effective in identifying very low levels of the organism. The three methods were used to detect the eae gene over a range of DNA concentrations. The differences in sensitivity were statistically significant (p < 0.05), and SYBR Green I PCR was found to have the lowest detection limit of the three; LUX primers had the highest detection limit. Therefore, using a defined DNA concentration for detecting the eae gene, SYBR Green I is the best alternative.

Clinical Validation of a Real-Time Polymerase Chain Reaction Detection of Neisseria gonorrheae porA Pseudogene Versus Culture Techniques

2008 ◽  
Vol 35 (5) ◽  
pp. 517-520 ◽  
Author(s):  
Stig Ove Hjelmevoll ◽  
Merethe Elise Olsen ◽  
Johanna U.Ericson Sollid ◽  
Håkon Haaheim ◽  
Kjetil K. Melby ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Clinical Validation ◽  
Polymerase Chain Reaction Detection ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Polymerase Chain

scholarly journals Multiplex Polymerase chain reaction to diagnose bloodstream infections in patients after cardiothoracic surgery

2018 ◽  
Author(s):  
Kevin Pilarczyk ◽  
Peter-Michael Rath ◽  
Jörg Steinmann ◽  
Matthias Thielmann ◽  
Maximillian Dürbeck ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Cardiac Surgery ◽  
Blood Culture ◽  
Real Time ◽  
Multiplex Polymerase Chain Reaction ◽  
Bloodstream Infections ◽  
Chain Reaction ◽  
Blood Samples ◽  
Gram Negative ◽  
Polymerase Chain

Abstract Background: Sepsis and other infectious complications are major causes of mortality and morbidity in patients after cardiac surgery. Whereas blood culture (BC) as the current diagnostic gold standard suffers from low sensitivity as well as a reporting delay of approximately 48–72 h, polymerase chain reaction (PCR) based technologies might offer a fast and reliable alternative for detection of bloodstream infections (BSI). The aim of this study was to compare the performance of real time multiplex-PCR “SeptiFast” (SF), a real-time multiplex PCR assay, with conventional BC testing in patients after cardiac surgery. Methods: 279 blood samples from 168 individuals with suspected BSI were analyzed by SF and BC. Receiver operating characteristic (ROC) curves were generated to determine the accuracy of clinical and laboratory information for the prediction of positive SF results. Results: Excluding results attributable to contaminants, 14.7% (n = 41) of blood samples were positive using SF and 17.2% (n=49) using conventional BC (p= n.s.). In six samples, SF detected more than one pathogen. Among the 47 microorganisms identified by SF, only 11 (23.4%) could be confirmed by BC. SF identified a significantly higher number of Gram-negative bacteria than BC (28 vs. 12, χ2=7.97, p=0.005). The combination of BC and SF significantly increased the number of detected microorganism, including fungi, when compared to BC alone (86 vs. 49, χ2=13.51, p<0.001). C-reactive protein (CRP) (21.7±11.41 vs. 16.0±16.9 mg/dl, p=0.009), procalcitonin (PCT) (28.7±70.9 vs. 11.5±30.4 ng/dl, p=0.015) as well as interleukin 6 (IL 6) (932.3±1306.7 vs. 313.3±686.6 pg/ml, p=0.010) was significantly higher in patients with a positive SF result. In addition, incidence of severe acute kidney injury (AKI) was higher in SF positive than in SF negative patients (31/42 [76%] vs. 125/237 [53%], p=0.01). Using ROC analysis, IL-6 (AUC 0.836) as well as CRP (AUC 0.804), but not PCT showed the best predictive values for positive SF results. Microbiological diagnostic information gained through SF led to 8 therapy adaptations. Conclusion: The real time PCR-based SF test might represent a valuable addition to the traditional BC method for rapid etiologic diagnosis of BSI in patients after cardiothoracic surgery. This powerful method furthermore applies in particular for individuals with fungal infections, Gram-negative bacteremia, AKI and/or elevated CRP and IL-6-concentration. However, due to the low performance in detecting Gram-positive pathogens and the inability to determine antibiotic susceptibility, it should always be used in combination with BC. [1] Key words: Blood stream infection, blood culture, real time multiplex Polymerase Chain Reaction

scholarly journals Real-Time Polymerase Chain Reaction for Microbiological Diagnosis of Parapneumonic Effusions in Canadian Children

2014 ◽  
Vol 25 (3) ◽  
pp. 151-154 ◽  
Author(s):  
Jeffrey M Pernica ◽  
Ioana Moldovan ◽  
Francis Chan ◽  
Robert Slinger
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Pleural Fluid ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Traditional Culture ◽  
Parapneumonic Effusion ◽  
Microbiological Diagnosis ◽  
Chain Reaction ◽  
Polymerase Chain

BACKGROUND: Community-acquired pneumonia (CAP) complicated by parapneumonic effusion/empyema is an infectious syndrome commonly encountered by physicians caring for children in Canada.OBJECTIVE: To investigate the incremental benefit of novel molecular testing for the microbiological diagnosis of pediatric CAP complicated by parapneumonic effusion/empyema in Canada.METHODS: A convenience sample of pleural fluid from 56 children who had been admitted to hospital in Ontario with CAP complicated by parapneumonic effusion between 2009 and 2011 was examined. Multiple uniplex real-time polymerase chain reaction (PCR) testing was performed on these pleural fluids and compared with traditional culture-based testing of blood and pleural fluid samples.RESULTS: Molecular methods detected a pathogen in 82% of cases, whereas traditional cultures of blood and pleural fluids detected a pathogen in only 25%. The majority of parapneumonic effusions were associated with pneumococcal infection;Streptococcus pneumoniaewas detected in 62% of the samples using molecular methods but in only 14% of samples using culture-based methods.Streptococcus pyogenes, detected in 16% of samples using PCR, was the second most common pathogen found. No patients were found to have empyema caused byStaphylococcus aureus.DISCUSSION: The results showed that multiple uniplex real-time PCR performed substantially better than traditional culture methods for microbiological diagnosis of CAP complicated by effusion/ empyema.S pneumoniaeandS pyogeneswere found to be responsible for the majority of infections. The approach detected pathogens in a similar proportion of pleural fluid samples as previously reported nested PCR assays; furthermore, the real-time closed-well approach also minimized the risk of nonspecificity due to cross-contamination relative to nested PCR.CONCLUSIONS: Real-time PCR for the detection of bacterial DNA in pleural fluids has the potential to better define the microbiological cause of pediatric CAP. This approach could help clinicians provide targeted antimicrobial therapy.

scholarly journals Cost Analysis of Real-Time Polymerase Chain Reaction Microbiological Diagnosis in Patients with Septic Shock

2012 ◽  
Vol 40 (6) ◽  
pp. 958-963 ◽  
Author(s):  
J. Alvarez ◽  
J. Mar ◽  
E. Varela-Ledo ◽  
M. Garea ◽  
L. Matinez-Lamas ◽  
...  
Keyword(s):  
Septic Shock ◽  
Polymerase Chain Reaction ◽  
Cost Analysis ◽  
Real Time ◽  
Microbiological Diagnosis ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Rapid antibiotic susceptibility testing from blood culture bottles with species agnostic real-time polymerase chain reaction

PLoS ONE ◽  
2018 ◽  
Vol 13 (12) ◽  
pp. e0209042 ◽  
Author(s):  
Tucker Maxson ◽  
Candace D. Blancett ◽  
Amanda S. Graham ◽  
Christopher P. Stefan ◽  
Timothy D. Minogue
Keyword(s):  
Polymerase Chain Reaction ◽  
Blood Culture ◽  
Real Time ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Antibiotic Susceptibility Testing ◽  
Chain Reaction ◽  
Polymerase Chain
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