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 ◽  
...  

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

2001 ◽  
Vol 13 (4) ◽  
pp. 308-311 ◽  
Author(s):  
Jacek Osek

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.


2014 ◽  
Vol 38 (1) ◽  
pp. 99-106
Author(s):  
Ihab G. M. AL-Shemmari

The aim of this study was to identify pasteurella multocida and their types by PCR in cattle’s and buffaloesi bagdad from March to August 2012 on 204 animals , including 102 cattle and 102 buffaloes at slaughter houses from Baghdad .Blood samples and nasal swaps were collected , before slaughtering and lung tissues of slaughtered animal , and from 54 clinically suspected cases of pasteurellosis , including 27 bovines ,and 27 buffaloes the samples taken included blood and nasal swabs . Pasteurellamultocida were isolated from 94 animals include 49 cattle 45 buffaloes. The typing of the isolates by multiplex PCR for genotyping Pasteuerllamultocida revealed 93 isolates of type B , 31 from cattle and 62 from buffaloes ,and 81 isolates of type A , 55 from cattle and 26 from buffaloes .


2015 ◽  
Vol 61 (7) ◽  
pp. 1071-1080 ◽  
Author(s):  
Ritu Banerjee ◽  
Christine B. Teng ◽  
Scott A. Cunningham ◽  
Sherry M. Ihde ◽  
James M. Steckelberg ◽  
...  

2010 ◽  
Vol 51 (9) ◽  
pp. 1042-1049 ◽  
Author(s):  
Massimo Resti ◽  
Maria Moriondo ◽  
Martina Cortimiglia ◽  
Giuseppe Indolfi ◽  
Clementina Canessa ◽  
...  

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