New developments in the diagnosis of bloodstream infections

Peters, Remco P. H.; Agtmael, Michiel A. van; Danner, Sven A.; Savelkoul, Paul H. M.; Vandenbroucke-Grauls, Christina M. J. E.

doi:10.1016/s1473-3099(04)01205-8

Cited by 357 publications

(320 citation statements)

References 96 publications

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“…Real-time PCR technology can detect minute amounts of pathogen DNA in patient blood samples with results available within hours [9]. Laboratory validation studies have focused on two approaches using PCR for genomic amplification with either (a) broad range detection of bacterial or fungal DNA with universal primers, followed by species identification using a post-PCR technique such as gene sequencing or electrospray mass spectrometry or (b) using species-specific hybridisation probes that provide direct confirmation of the species present [10].…”

Section: Introductionmentioning

confidence: 99%

Accuracy of LightCycler® SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review and meta-analysis

et al. 2014

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2 AbstractPurpose. There is an urgent need to develop diagnostic tests to improve the detection of pathogens causing life-threatening infection (sepsis). SeptiFast is a CEmarked multi-pathogen real-time PCR system capable of detecting DNA sequences of bacteria and fungi present in blood samples within a few hours. We report here a systematic review and meta-analysis of diagnostic accuracy studies of SeptiFast in the setting of suspected sepsis.Methods. A comprehensive search strategy was developed to identify studies that compared SeptiFast with blood culture in suspected sepsis. Methodological quality was assessed using QUADAS. Heterogeneity of studies was investigated using a coupled forest plot of sensitivity and specificity and a scatter plot in Receiver Operator Characteristic space. Bivariate model method was used to estimate summary sensitivity and specificity.Results. From 41 phase III diagnostic accuracy studies, summary sensitivity and specificity for SeptiFast compared with blood culture were 0.68 (95% CI 0.63-0.73) and 0.86 (95% CI 0.84-0.89) respectively. Study quality was judged to be variable with important deficiencies overall in design and reporting that could impact on derived diagnostic accuracy metrics.Conclusions. SeptiFast appears to have higher specificity than sensitivity, but deficiencies in study quality are likely to render this body of work unreliable. Based on the evidence presented here, it remains difficult to make firm recommendations about the likely clinical utility of SeptiFast in the setting of suspected sepsis. We recommend that future studies should include well designed and reported clinical diagnostic accuracy elements measured against all of the features of the STARD criteria to help inform the subsequent design of much needed interventional studies in the management of suspected sepsis. Word count: 225 2413

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Section: Introductionmentioning

confidence: 99%

Accuracy of LightCycler® SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review and meta-analysis

et al. 2014

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“…In the case of sepsis, culture is considered the mainstay for microbiological diagnosis, but it often does not provide timecritical results for optimal early management (Bauer & Reinhart, 2010;Jorgensen et al, 1997) and lacks sensitivity in patients already treated with antibiotics (Peters et al, 2004). The LightCycler SeptiFast (SF) test (Roche Diagnostics) is a commercially available multiplex real-time PCR assay able to detect and differentiate a wide range of Gram-negative and Gram-positive bacteria, and some fungi, commonly involved in systemic infections (Lehmann et al, 2008) (shown in Table 1).…”

Section: Introductionmentioning

confidence: 99%

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood

Mencacci

Leli

Cardaccia

et al. 2011

Journal of Medical Microbiology

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Early detection of aetiological agents is pivotal for adequate therapy for bacterial infections. Although culture is still considered the mainstay for laboratory diagnosis, it often lacks sensitivity, especially in patients already treated with antibiotics. The present study investigated the potential clinical utility of the commercial real-time-PCR-based system SeptiFast (SF), originally intended for diagnosis of sepsis from blood specimens, in the aetiological diagnosis of other bacterial infections, in patients undergoing antibiotic therapy. A total of 53 non-blood specimens were analysed for microbial pathogen detection by conventional culture and with SF real-time PCR: 19 (35.8 %) synovial fluids, 9 (17.0 %) cardiac valve tissues and 25 (47.2 %) purulent exudates from various body sites. Overall, the number of specimens positive for a pathogen by SF (26/53; 49.1 %) was significantly greater (P50.001) than that of specimens positive by culture (10/53; 18.9 %). In particular, SF was superior to culture for pathogen detection in cardiac valve tissues and synovial fluids. The analysis of concordance showed a fair agreement between the two methods (kappa value50.314; 95 % confidence interval50.531-0.097). Even with the limitation of the low number of specimens, this study confirmed the great potential of diagnosing bacterial infections by a molecular approach, and indicates that the real-time PCR SF system can be used for specimens other than blood, from patients undergoing antibiotic treatment.

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“…Fast and accurate diagnosis is important for reduction of the mortality of LOS. Although blood culture remains to be the gold standard in the diagnosis of bacterial bloodstream infections, this method has certain limitations, such as a long waiting time for results (at least 48 h), poor sensitivity (10-20%) in detecting fastidious microbes, and the use of antibiotics before blood specimens are drawn, which may affect the results obtained from the blood culture (4,5). In addition, the blood culture method is flawed in detection of polymicrobial infection (5,6).…”

Section: Introductionmentioning

confidence: 99%

16S rDNA PCR-DGGE and sequencing in the diagnosis of neonatal late-onset septicemia

Liu

et al. 2015

Molecular Medicine Reports

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Abstract. The 16S rDNA polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and sequencing method has been demonstrated to be valuable in detecting pathogens in the blood of patients suffering from fever or neutropenia. However, its use in the diagnosis of neonatal late-onset septicemia (LOS) has not yet been reported. The aim of the present study was to investigate the efficiency of this method in detection of the type of bacterial infection in neonatal LOS. Blood specimens from 60 neonates in whom LOS was suspected were collected. Fourteen culture positive blood samples and 24 spiked 'infected' blood samples were analyzed by the 16S rDNA PCR-DGGE and sequencing method or by pathogen-specific PCR. Only in 5 of the 14 cases did the results of 16S rDNA PCR-DGGE and sequencing match with the results of blood culturing. In the other 9 cases, the blood culture failed to detect bacteria, such as Neisseria sp. and Moraxella sp., which were detected by 16S rDNA PCR-DGGE and sequencing. Furthermore, the 16S rDNA PCR-DGGE and sequencing failed to detect blood culture-proven bacteria, such as Klebsiella pneumonia. A competitive inhibitory effect in 16S rDNA PCR amplification may lead to the discrepancy between pathogen-specific PCR and spiked 'infected' blood samples. When a certain species of bacteria was detected by 16S rDNA PCR, the competitive inhibitory effect presented a higher sensitivity in detecting this species in the blood samples that contained bacterial DNA only from this species compared with the blood samples that were blended with other bacterial DNAs. In conclusion, 16S rDNA PCR-DGGE and sequencing can detect a more comprehensive spectrum of pathogens than blood culture. However, the competitive inhibitory effect, which may lead to false negative results should be taken into consideration when the 16S rDNA PCR-DGGE and sequencing method is applied to the diagnosis of neonatal LOS.

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New developments in the diagnosis of bloodstream infections

Cited by 357 publications

References 96 publications

Accuracy of LightCycler® SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review and meta-analysis

Accuracy of LightCycler® SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review and meta-analysis

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood

16S rDNA PCR-DGGE and sequencing in the diagnosis of neonatal late-onset septicemia

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