Abstract
Purpose
There is an urgent need to develop diagnostic tests to improve the detection of pathogens causing life-threatening infection (sepsis). SeptiFast is a CE-marked 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.
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References
Bone RC, Sibbald WJ, Sprung CL (1992) The ACCP-SCCM consensus conference on sepsis and organ failure. Chest 101:1481–1483
Dellinger RP, Levy MM, Rhodes A, Annane D, Gerlach H, Opal SM, Sevransky JE, Sprung CL, Douglas IS, Jaeschke R, Osborn TM, Nunnally ME, Townsend SR, Reinhart K, Kleinpell RM, Angus DC, Deutschman CS, Machado FR, Rubenfeld GD, Webb S, Beale RJ, Vincent JL, Moreno R (2012) Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock. Intensive Care Med 39:165–228
Calandra T, Cohen J (2005) The international sepsis forum consensus conference on definitions of infection in the intensive care unit. Crit Care Med 33:1538–1548
Health Protection Agency (2005) Investigation of blood cultures (for organisms other than mycobacterium species). National Standard Method, Standards Unit, London
Comptroller Auditor General (2009) Reducing healthcare associated infection in hospitals in England. HC 560 Session 2008–2009. The Stationery Office, London
Kollef MH, Sherman G, Ward S, Fraser VJ (1999) Inadequate antimicrobial treatment of infections: a risk factor for hospital mortality among critically ill patients. Chest 115:462–474
Kumar A, Roberts D, Wood KE, Light B, Parrillo JE, Sharma S, Suppes R, Feinstein D, Zanotti S, Taiberg L, Gurka D, Kumar A, Cheang M (2006) Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 34:1589–1596
World Health Organisation (2012) The evolving threat of antimicrobial resistance: options for action. World Health Organisation, Geneva
Peters RP, van Agtmael MA, Danner SA, Savelkoul PH, Vandenbroucke-Grauls CM (2004) New developments in the diagnosis of bloodstream infections. Lancet Infect Dis 4:751–760
Dark PM, Dean P, Warhurst G (2009) Bench-to-bedside review: the promise of rapid infection diagnosis during sepsis using polymerase chain reaction-based pathogen detection. Crit Care 13:217
Lehmann LE, Hunfeld KP, Emrich T, Haberhausen G, Wissing H, Hoeft A, Stuber F (2008) A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples. Med Microbiol Immunol 197:313–324
Chang SS, Hsieh WH, Liu TS, Lee SH, Wang CH, Chou HC, Yeo YH, Tseng CP, Lee CC (2013) Multiplex PCR system for rapid detection of pathogens in patients with presumed sepsis—a systemic review and meta-analysis. PLoS One 8:e62323
Dark PM, Dunn G, Chadwick P, Young D, Bentley A, Carlson G, Warhurst G (2011) The clinical diagnostic accuracy of rapid detection of healthcare-associated bloodstream infection in intensive care using multi-pathogen real-time polymerase chain reaction (real-time PCR) technology. BMJ Open 1:e000181
Dark P, Wilson C, Blackwood B, McAuley DF, Perkins GD, McMullan R, Gates S, Warhurst G (2011) Accuracy of LightCycler SeptiFast for the detection and identification of pathogens in blood of patients with suspected sepsis: a systematic review. PROSPERO CRD42011001289. http://www.crd.york.ac.uk/PROSPERO/full_doc.asp?ID=CRD42011001289
Dark P, Wilson C, Blackwood B, McAuley DF, Perkins GD, McMullan R, Gates S, Warhurst G (2012) Accuracy of LightCycler(R) SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review protocol. BMJ Open 2:e000392
Levy MM, Fink MP, Marshall JC, Abraham E, Angus D, Cook D, Cohen J, Opal SM, Vincent JL, Ramsay G (2003) 2001 SCCM/ESICM/ACCP/ATS/SIS International Sepsis Definitions Conference. Intensive Care Med 29:530–538
Whiting P, Rutjes AW, Reitsma JB, Bossuyt PM, Kleijnen J (2003) The development of QUADAS: a tool for the quality assessment of studies of diagnostic accuracy included in systematic reviews. BMC Med Res Methodol 3:25
Reitsma JB, Glas AS, Rutjes AW, Scholten RJ, Bossuyt PM, Zwinderman AH (2005) Bivariate analysis of sensitivity and specificity produces informative summary measures in diagnostic reviews. J Clin Epidemiol 58(10):982–990
Harbord RM, Deeks JJ, Egger M, Whiting P, Sterne JA (2007) A unification of models for meta-analysis of diagnostic accuracy studies. Biostatistics 8:239–251
Phillips B, Ball C, Sackett D, Badenoch D, Straus S, Haynes B, Dawes M (2001) Oxford Centre for Evidence-based Medicine. Levels of evidence. http://www.cebm.net/index.aspx?O=1025. Accessed 6 Jan 2012
Raglio A, Rizzi M, Amer M, Mangia M, Lucà M, Goglio A (2006) Sepsis diagnosis by realtime PCR (SeptiFast Kit, Roche Diagnostics). In: Abstracts of the 16th European Congress of Clinical Microbiology and Infectious Diseases, Nice, France, April 1–4 2006. Blackwell, Oxford, O215
Klemm M, Prinz M, Nowak A, Morgenstern T, Meisner M, Rothe KF, Demant T (2007) Clinical application of SeptiFast, a PCR method for the detection of bacteraemia, in intensive care patients. Infection 35:12
Bingold TM, Hunfeld K, Scheller B, Rönneberg T, Sartorius S, Wahrmann M, Klösel S, Zwißler B, Wissing H (2007) Clinical utility of a new PCR-based assay for rapid pathogen detection (SeptiFast®) in patients with clinical sepsis compared to standard blood culture. Infection 35:15
Vince A, Lepej SZ, Barsic B, Dusek D, Mitrovic Z, Serventi-Seiwerth R, Labar B (2008) LightCycler SeptiFast assay as a tool for the rapid diagnosis of sepsis in patients during antimicrobial therapy. J Med Microbiol 57:1306–1307
Mancini N, Clerici D, Diotti R, Perotti M, Ghidoli N, De Marco D, Pizzorno B, Emrich T, Burioni R, Ciceri F, Clementi M (2008) Molecular diagnosis of sepsis in neutropenic patients with haematological malignancies. J Med Microbiol 57:601–604
Louie RF, Tang Z, Albertson TE, Cohen S, Tran NK, Kost GJ (2008) Multiplex polymerase chain reaction detection enhancement of bacteremia and fungemia. Crit Care Med 36:1487–1492
Lodes U, Bohmeier B, Meyer F, Koenig B, Lippert H (2008) Microbiologic diagnostic with the novel Lightcycler Septifast® test during the early phase of surgical sepsis mainly caused by peritonitis is quicker and more sensitive than conventional microbiological culture. Gastroenterology 134:A621
Westh H, Lisby G, Breysse F, Boddinghaus B, Chomarat M, Gant V, Goglio A, Raglio A, Schuster H, Stuber F, Wissing H, Hoeft A (2009) Multiplex real-time PCR and blood culture for identification of bloodstream pathogens in patients with suspected sepsis. Clin Microbiol Infect 15:544–551
Varani S, Stanzani M, Paolucci M, Melchionda F, Castellani G, Nardi L, Landini MP, Baccarani M, Pession A, Sambri V (2009) Diagnosis of bloodstream infections in immunocompromised patients by real-time PCR. J Infect 58:346–351
Palomares J, Puche B, Martos A, Lucena F, Marín M, Martín-Mazuelos E (2009) Rapid molecular diagnosis of severe sepsis in patients with symptoms of severe sepsis. In: Abstracts of the 19th European Congress of Clinical Microbiology and Infectious Diseases, Helsiniki, Finland, 16–19 May 2009. Blackwell, Oxford, p 1827
Lodes U, Meyer F, Konig B, Lippert H (2009) Microbiological sepsis screening in surgical ICU patients with the “lightCycler” Septifast test—a pilot study. Zentralbl Chir 134:249–253
Dierkes C, Ehrenstein B, Siebig S, Linde HJ, Reischl U, Salzberger B (2009) Clinical impact of a commercially available multiplex PCR system for rapid detection of pathogens in patients with presumed sepsis. BMC Infect Dis 9:126
Dark P, Chadwick P, Warhurst G (2009) Detecting sepsis-associated bloodstream infection acquired in intensive care using multi-pathogen real-time PCR. J Infect 59:296–298
Yanagihara K, Kitagawa Y, Tomonaga M, Tsukasaki K, Kohno S, Seki M, Sugimoto H, Shimazu T, Tasaki O, Matsushima A, Ikeda Y, Okamoto S, Aikawa N, Hori S, Obara H, Ishizaka A, Hasegawa N, Takeda J, Kamihira S, Sugahara K, Asari S, Murata M, Kobayashi Y, Ginba H, Sumiyama Y, Kitajima M (2010) Evaluation of pathogen detection from clinical samples by real-time polymerase chain reaction using a sepsis pathogen DNA detection kit. Crit Care 14:R159
Wallet F, Nseir S, Baumann L, Herwegh S, Sendid B, Boulo M, Roussel-Delvallez M, Durocher AV, Courcol RJ (2010) Preliminary clinical study using a multiplex real-time PCR test for the detection of bacterial and fungal DNA directly in blood. Clin Microbiol Infect 16:774–779
Tsalik EL, Jones D, Nicholson B, Waring L, Liesenfeld O, Park LP, Glickman SW, Caram LB, Langley RJ, van Velkinburgh JC, Cairns CB, Rivers EP, Otero RM, Kingsmore SF, Lalani T, Fowler VG, Woods CW (2010) Multiplex PCR to diagnose bloodstream infections in patients admitted from the emergency department with sepsis. J Clin Microbiol 48:26–33
Soki J, Szabó E, Hajdú E, Lázár A, Nagy E (2010) Use of a commercially available multiplex real-time PCR assay (SeptiFast) to detect bacterial and fungal pathogens in septic patients. In: 20th European Congress of Clinical Microbiology and Infectious Diseases, 2010, Vienna, Austria. European Congress of Clinical Microbiology and Infectious Diseases
Regueiro BJ, Varela-Ledo E, Martinez-Lamas L, Rodriguez-Calvino J, Aguilera A, Santos A, Gomez-Tato A, Alvarez-Escudero J (2010) Automated extraction improves multiplex molecular detection of infection in septic patients. PLoS One 5:e13387
Maubon D, Hamidfar-Roy R, Courby S, Vesin A, Maurin M, Pavese P, Ravanel N, Bulabois CE, Brion JP, Pelloux H, Timsit JF (2010) Therapeutic impact and diagnostic performance of multiplex PCR in patients with malignancies and suspected sepsis. J Infect 61:335–342
Lehmann LE, Hunfeld KP, Steinbrucker M, Brade V, Book M, Seifert H, Bingold T, Hoeft A, Wissing H, Stuber F (2010) Improved detection of blood stream pathogens by real-time PCR in severe sepsis. Intensive Care Med 36:49–56
Bloos F, Hinder F, Becker K, Sachse S, Mekontso Dessap A, Straube E, Cattoir V, Brun-Buisson C, Reinhart K, Peters G, Bauer M (2010) A multicenter trial to compare blood culture with polymerase chain reaction in severe human sepsis. Intensive Care Med 36:241–247
Berger A, Altiok I, Mechtler T, Langgartner M, Böhm J, Herkner K, Pollack A, Kasper D (2010) Adaption of the Roche SeptiFast system for the early detection of neonatal sepsis in very low birthweight infants. Klinische Pädiatrie 222:S4
Avolio M, Diamante P, Zamparo S, Modolo ML, Grosso S, Zigante P, Tosoni N, De Rosa R, Stano P, Camporese A (2010) Molecular identification of bloodstream pathogens in patients presenting to the emergency department with suspected sepsis. Shock 34:27–30
Vrioni G, Daniil I, Mamali V, Kimouli M, Mylona-Petropoulou D, Themeli-Digalaki K, Tsakris A (2011) Preliminary clinical study using a multiplex blood PCR for rapid detection of bacterial and fungal pathogens in ICU patients with presumed sepsis. In: 21st European Congress of Clinical Microbiology and Infectious Diseases, 2011, Milan, Italy. European Congress of Clinical Microbiology and Infectious Diseases
Sitnik R, Marra AR, Petroni RC, Ramos OP, Martino MD, Pasternak J, Santos OS, Mangueira CL, Rebello Pinho JR (2011) SeptiFast for diagnosis of sepsis in severely ill patients from a Brazilian Hospital. J Med Diagn 13:736
Obara H, Aikawa N, Hasegawa N, Hori S, Ikeda Y, Kobayashi Y, Murata M, Okamoto S, Takeda J, Tanabe M, Sakakura Y, Ginba H, Kitajima M, Kitagawa Y (2011) The role of a real-time PCR technology for rapid detection and identification of bacterial and fungal pathogens in whole-blood samples. J Infect Chemother 17:327–333
Lucignano B, Ranno S, Liesenfeld O, Pizzorno B, Putignani L, Bernaschi P, Menichella D (2011) Multiplex PCR allows rapid and accurate diagnosis of bloodstream infections in newborns and children with suspected sepsis. J Clin Microbiol 49:2252–2258
Josefson P, Stralin K, Ohlin A, Ennefors T, Dragsten B, Andersson L, Fredlund H, Molling P, Olcen P (2011) Evaluation of a commercial multiplex PCR test (SeptiFast) in the etiological diagnosis of community-onset bloodstream infections. Eur J Clin Microbiol Infect Dis 30:1127–1134
Hettwer S, Wilhelm J, Schurmann M, Ebelt H, Hammer D, Amoury M, Hofmann F, Oehme A, Wilhelms D, Kekule AS, Kloss T, Werdan K (2011) Microbial diagnostics in patients with presumed severe infection in the emergency department. Med Klin Intensivmed Notfmed 107:53–62
Bravo D, Blanquer J, Tormo M, Aguilar G, Borras R, Solano C, Clari MA, Costa E, Munoz-Cobo B, Argueso M, Pineda JR, Navarro D (2011) Diagnostic accuracy and potential clinical value of the LightCycler SeptiFast assay in the management of bloodstream infections occurring in neutropenic and critically ill patients. Int J Infect Dis 15:e326–e331
Tschiedel E, Steinmann J, Buer J, Onnebrink JG, Felderhoff-Muser U, Rath PM, Dohna-Schwake C (2012) Results and relevance of molecular detection of pathogens by SeptiFast–a retrospective analysis in 75 critically ill children. Klin Padiatr 224:12–16
Rath PM, Saner F, Paul A, Lehmann N, Steinmann E, Buer J, Steinmann J (2012) Multiplex PCR for rapid and improved diagnosis of bloodstream infections in liver transplant recipients. J Clin Microbiol 50:2069–2071
Pasqualini L, Mencacci A, Leli C, Montagna P, Cardaccia A, Cenci E, Montecarlo I, Pirro M, di Filippo F, Cistaro E, Schillaci G, Bistoni F, Mannarino E (2012) Diagnostic performance of a multiple real-time PCR assay in patients with suspected sepsis hospitalized in an internal medicine ward. J Clin Microbiol 50:1285–1288
Mauro MV, Cavalcanti P, Perugini D, Noto A, Sperli D, Giraldi C (2012) Diagnostic utility of LightCycler SeptiFast and procalcitonin assays in the diagnosis of bloodstream infection in immunocompromised patients. Diagn Microbiol Infect Dis 73:308–311
Lodes U, Bohmeier B, Lippert H, Konig B, Meyer F (2012) PCR-based rapid sepsis diagnosis effectively guides clinical treatment in patients with new onset of SIRS. Langenbecks Arch Surg 397:447–455
Guido M, Quattrocchi M, Zizza A, Pasanisi G, Pavone V, Lobreglio G, Gabutti G, De Donno A (2012) Molecular approaches in the diagnosis of sepsis in neutropenic patients with haematological malignances. J Prev Med Hyg 53:104–108
Grif K, Fille M, Wurzner R, Weiss G, Lorenz I, Gruber G, Eschertzhuber S, Nachbaur D, Lass-Florl C, Orth D (2012) Rapid detection of bloodstream pathogens by real-time PCR in patients with sepsis. Wien Klin Wochenschr 124:266–270
Leitner E, Kessler HH, Spindelboeck W, Hoenigl M, Putz-Bankuti C, Stadlbauer-Kollner V, Krause R, Grisold AJ, Feierl G, Stauber RE (2013) Comparison of two molecular assays with conventional blood culture for diagnosis of sepsis. J Microbiol Methods 3:253–255
Herne V, Nelovkov A, Kütt M, Ivanova M (2013) Diagnostic performance and therapeutic impact of LightCycler SeptiFast assays in patients with suspected sepsis. Eur J Microbiol Immunol 3:68–76
Burdino E, Ruggiero T, Allice T, Milia MG, Gregori G, Milano R, Cerutti F, De Rosa FG, Manno E, Caramello P, Di Perri G, Ghisetti V (2014) Combination of blood cultures and SeptiFast molecular test in aptients with suspected sepsis for the identification of bloodstream pathogens. Diagn Microbiol Infect Dis 79:287–292
Avolio M, Diamante P, Modolo ML, DeRosa R, Stano P, Camporese A (2014) Direct molecular detection of pathogens in blood as specific rule-in diagnsotic biomarker in patients with presumed sepsis: our experince on a heterogeneous cohort of patients with signs of infective systemic inflammatory response syndrome. Shock 42:86–92
Bossuyt PM, Reitsma JB, Bruns DE, Gatsonis CA, Glasziou PP, Irwig LM, Lijmer JG, Moher D, Rennie D, de Vet HC (2003) Towards complete and accurate reporting of studies of diagnostic accuracy: the STARD initiative. BMJ 326:41–44
Rutjes AWS, Reitsma JB, Coomarasamy A, Khan KS, Bossuyt PMM (2007) Evaluation of diagnostic tests when there is no gold standard. A review of methods. Health Technol Assess 11(50):iii, ix–51
Acknowledgements
We thank Mrs Helen McEvoy, formerly at the University of Manchester, School of Medicine Library, for her assistance with search strategy design and Mrs Valerie Haigh, Salford Royal NHS Foundation Trust Library and Information Resource Centre, for her advice on the use of filters in diagnostic search strategies. PD and GW were part-funded by the UK Health Technology Assessment (HTA) programme of the National Institute of Health Research grant number NIHR HTA 08/13/16. CW and DG were funded by NIHR Greater Manchester Comprehensive Local Research Network in support of this programme of work. DFM and GDP receive funding from The Intensive Care Foundation (UK) in support of their roles as National Research Directors. None of these funding organisations or any commercial organisation have contributed to the study design; collection, management, analysis and interpretation of data; writing of the report or the decision to submit the report for publication. Only the named authors have ultimate authority over each of these activities.
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Dark, P., Blackwood, B., Gates, S. et al. 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. Intensive Care Med 41, 21–33 (2015). https://doi.org/10.1007/s00134-014-3553-8
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DOI: https://doi.org/10.1007/s00134-014-3553-8