A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples

Lehmann, Lutz; Hunfeld, Klaus-Peter; Emrich, Thomas; Haberhausen, Gerd; Wissing, H.; Hoeft, Andreas; Stüber, Frank

doi:10.1007/s00430-007-0063-0

Cited by 280 publications

(313 citation statements)

References 30 publications

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“…SeptiFast has been assessed at the laboratory level on clinical isolates and shown to have good analytical specificity and exclusivity, confirming its analytical validity [11]. Numerous clinical diagnostic studies have been reported since SeptiFast gained its CE-mark in 2006, predominantly focused on suspected sepsis.…”

Section: Introductionmentioning

confidence: 83%

“…LightCycler® SeptiFast as the index test on blood specimens for the detection and simultaneous identification of bacterial and fungal pathogens [11].…”

Section: Index Testmentioning

confidence: 99%

“…SeptiFast, manufactured by Roche Diagnostics, GmbH, Mannheim, Germany and run on their real-time PCR instrument (the LightCycler®), was the first real-time PCR based system to be awarded a Conformité Européenne (CE) mark for simultaneous pathogen detection and identification in suspected bloodstream infection [11]. The system uses a multiplex approach, which allows detection of the most common pathogen species causing bloodstream infection in a single blood sample (Table 1).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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: 83%

“…LightCycler® SeptiFast as the index test on blood specimens for the detection and simultaneous identification of bacterial and fungal pathogens [11].…”

Section: Index Testmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…The IPC amplification demonstrates the absence of PCR inhibitors. Therefore, the concomitant absence of sample amplification can be considered as a valid result [27].…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…[21,27,43]. PCR is the most widely used molecular diagnostic technique due to its fast and easy to use protocol.…”

Section: Pcr Implementation In Diagnosticsmentioning

confidence: 99%

Potentials and limitations of molecular diagnostic methods in food safety

Lauri

Mariani

2008

Genes Nutr

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Molecular methods allow the detection of pathogen nucleic acids (DNA and RNA) and, therefore, the detection of contamination in food is carried out with high selectivity and rapidity. In the last 2 decades molecular methods have accompanied traditional diagnostic methods in routine pathogen detection, and might replace them in the upcoming future. In this review the implementation in diagnostics of four of the most used molecular techniques (PCR, NASBA, microarray, LDR) are described and compared, highlighting advantages and limitations of each of them. Drawbacks of molecular methods with regard to traditional ones and the difficulties encountered in pathogen detection from food or clinical specimen are also discussed. Moreover, criteria for the choice of the target sequence for a secure detection and classification of pathogens and possible developments in molecular diagnostics are also proposed.

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Polymerase chain reaction blood tests for the diagnosis of invasive aspergillosis in immunocompromised people

Cruciani

Mengoli

Loeffler³

et al. 2015

Cochrane Database of Systematic Reviews

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A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples

Cited by 280 publications

References 30 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

Potentials and limitations of molecular diagnostic methods in food safety

Polymerase chain reaction blood tests for the diagnosis of invasive aspergillosis in immunocompromised people

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