DNAemia Detection by Multiplex PCR and Biomarkers for Infection in Systemic Inflammatory Response Syndrome Patients

Fitting, Catherine; Parlato, Marianna; Adib‐Conquy, Minou; Mémain, N.; Philippart, François; Misset, Benoît; Monchi, Mehran; Cavaillon, Jean‐Marc; Adrie, Christophe

doi:10.1371/journal.pone.0038916

Cited by 70 publications

(49 citation statements)

References 4 publications

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“…Plasma samples from 20 patients (11 males/9 females, 64.4 ± 17.9 years, body mass index 25.0 ± 8.2 kg/m 2 ) with infectious SIRS but presenting a negative LAL test had been collected in a previous study ( 2 ). Sepsis in all patients was defi ned according to the criteria of the panel of experts from the American College of Chest Physician/Society of Critical Care Medicine ( 13 ).…”

Section: Human Plasma Samplesmentioning

confidence: 99%

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Barros

Gautier

Sali

et al. 2015

Journal of Lipid Research

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103

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Systemic infl ammatory response syndrome (SIRS) is defi ned by a cluster of clinical signs that include tachycardia, leukocytosis, tachypnoea, and pyrexia ( 1 ). Although SIRS is a major consequence of sepsis (i.e., a deleterious, nonresolving infl ammatory response to infection that can lead to multiple organ dysfunction and septic shock), it can be caused by many pathological events that are not associated with the bacterial infection ( 1 ). It is of major importance to distinguish between noninfectious SIRS, sepsis, and septic shock in critically ill patients because these groups of patients are markedly different in terms of care and clinical outcome. Positive microbiological identifi cation tests [using conventional microbiological tests or bacterial DNA detection ( 2 ) as well as a number of infl ammatory and infectious biomarkers, including cytokines, procalcitonin, immune cell markers or a combination of these ( 3 )] have been proposed, but at this stage, none has proved to be reliable enough to ascertain the occurrence and extent of infection in high-risk patients with SIRS. Interestingly, and as documented further by our group in the prospective multicenter cohort EPISS study ( 4 ), Gramnegative bacilli are the most frequently identifi ed pathogens in patients with septic shock. In this context, the direct quantitation of the culprit component of the Gram-negative bacterium [i.e., lipopolysaccharide (LPS), which triggers SIRS by interacting with the CD14/Toll-like receptor 4 /myeloid differentiation factor 2 receptor complex at the surface of leukocytes ( 1 ) Abbreviations: 3HM, 3-hydroxymyristic acid or 3-hydroxymyristate; LAL, limulus amebocyte lysate; LOD, limit of detection; LOQ, limit of quantifi cation; LPS, lipopolysaccharide; PLTP, phospholipid transfer protein; SIRS, systemic infl ammatory response syndrome; SOFA, sequential organ failure assessment .

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Section: Human Plasma Samplesmentioning

confidence: 99%

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Barros

Gautier

Sali

et al. 2015

Journal of Lipid Research

Self Cite

106

103

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“…Pathogens are detected by a 16S rRNA gene multiplex PCR. The overall turnaround time is approximately 8 h. In a study on 72 patients, concordance of a microbiological workup (mainly blood culture) and VYOO® test (VO) from whole blood was 46.2% [21]. VO was not able to detect clinical relevant pathogens in eight cases.…”

Section: Vyoo® Assaymentioning

confidence: 98%

Non-culture-based methods to diagnose bloodstream infection: Does it work?

Skvarč¹,

Stubljar²,

Rogina³

et al. 2013

European Journal of Microbiology and Immunology

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“…26,37 VYOO ® , introduced by SIRS-Lab, Jena, Germany, gained a CE mark in late 2008 and takes a multiplex approach to detect a panel of sepsis pathogens by electrophoretic separation of species-specific PCR amplicons. 38,39 PLEX-ID™ (Abbott, Wiesbaden, Germany) also uses universal PCR to detect a very broad range of bacterial and fungal pathogens, but utilises electrospray ionisation mass spectrometry for post-PCR species identification. 40 …”

Section: Background and Rationalementioning

confidence: 99%

Rapid detection of health-care-associated bloodstream infection in critical care using multipathogen real-time polymerase chain reaction technology: a diagnostic accuracy study and systematic review

Warhurst

Dunn

Chadwick

et al. 2015

Health Technology Assessment

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BackgroundThere is growing interest in the potential utility of real-time polymerase chain reaction (PCR) in diagnosing bloodstream infection by detecting pathogen deoxyribonucleic acid (DNA) in blood samples within a few hours. SeptiFast (Roche Diagnostics GmBH, Mannheim, Germany) is a multipathogen probe-based system targeting ribosomal DNA sequences of bacteria and fungi. It detects and identifies the commonest pathogens causing bloodstream infection. As background to this study, we report a systematic review of Phase III diagnostic accuracy studies of SeptiFast, which reveals uncertainty about its likely clinical utility based on widespread evidence of deficiencies in study design and reporting with a high risk of bias.ObjectiveDetermine the accuracy of SeptiFast real-time PCR for the detection of health-care-associated bloodstream infection, against standard microbiological culture.DesignProspective multicentre Phase III clinical diagnostic accuracy study using the standards for the reporting of diagnostic accuracy studies criteria.SettingCritical care departments within NHS hospitals in the north-west of England.ParticipantsAdult patients requiring blood culture (BC) when developing new signs of systemic inflammation.Main outcome measuresSeptiFast real-time PCR results at species/genus level compared with microbiological culture in association with independent adjudication of infection. Metrics of diagnostic accuracy were derived including sensitivity, specificity, likelihood ratios and predictive values, with their 95% confidence intervals (CIs). Latent class analysis was used to explore the diagnostic performance of culture as a reference standard.ResultsOf 1006 new patient episodes of systemic inflammation in 853 patients, 922 (92%) met the inclusion criteria and provided sufficient information for analysis. Index test assay failure occurred on 69 (7%) occasions. Adult patients had been exposed to a median of 8 days (interquartile range 4–16 days) of hospital care, had high levels of organ support activities and recent antibiotic exposure. SeptiFast real-time PCR, when compared with culture-proven bloodstream infection at species/genus level, had better specificity (85.8%, 95% CI 83.3% to 88.1%) than sensitivity (50%, 95% CI 39.1% to 60.8%). When compared with pooled diagnostic metrics derived from our systematic review, our clinical study revealed lower test accuracy of SeptiFast real-time PCR, mainly as a result of low diagnostic sensitivity. There was a low prevalence of BC-proven pathogens in these patients (9.2%, 95% CI 7.4% to 11.2%) such that the post-test probabilities of both a positive (26.3%, 95% CI 19.8% to 33.7%) and a negative SeptiFast test (5.6%, 95% CI 4.1% to 7.4%) indicate the potential limitations of this technology in the diagnosis of bloodstream infection. However, latent class analysis indicates that BC has a low sensitivity, questioning its relevance as a reference test in this setting. Using this analysis approach, the sensitivity of the SeptiFast test was low but also appeared significantly better than BC. Blood samples identified as positive by either culture or SeptiFast real-time PCR were associated with a high probability (> 95%) of infection, indicating higher diagnostic rule-in utility than was apparent using conventional analyses of diagnostic accuracy.ConclusionSeptiFast real-time PCR on blood samples may have rapid rule-in utility for the diagnosis of health-care-associated bloodstream infection but the lack of sensitivity is a significant limiting factor. Innovations aimed at improved diagnostic sensitivity of real-time PCR in this setting are urgently required. Future work recommendations include technology developments to improve the efficiency of pathogen DNA extraction and the capacity to detect a much broader range of pathogens and drug resistance genes and the application of new statistical approaches able to more reliably assess test performance in situation where the reference standard (e.g. blood culture in the setting of high antimicrobial use) is prone to error.Study registrationThe systematic review is registered as PROSPERO CRD42011001289.FundingThe National Institute for Health Research Health Technology Assessment programme. Professor Daniel McAuley and Professor Gavin D Perkins contributed to the systematic review through their funded roles as codirectors of the Intensive Care Foundation (UK).

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DNAemia Detection by Multiplex PCR and Biomarkers for Infection in Systemic Inflammatory Response Syndrome Patients

Cited by 70 publications

References 4 publications

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Non-culture-based methods to diagnose bloodstream infection: Does it work?

Rapid detection of health-care-associated bloodstream infection in critical care using multipathogen real-time polymerase chain reaction technology: a diagnostic accuracy study and systematic review

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