Adjuvant role of SeptiFast to improve the diagnosis of sepsis in a large cohort of hematological patients

Published online:


Febrile neutropenia and sepsis are common and life-threatening complications in hematological diseases. This study was performed retrospectively in 514 patients treated for febrile neutropenia at our institute, to investigate the clinical usefulness of a molecular tool, LightCycler® SeptiFast test (SF), to promptly recognize pathogens causing sepsis in hematological patients. We collected 1837 blood samples of 514 consecutive hematological patients. The time of processing is short. Overall, 757 microorganisms in 663 episodes were detected by molecular test and standard blood cultures (BC): 73.6% Gram-positive bacteria, 23.9% Gram-negative bacteria, and 2.5% fungal species. This large analysis demonstrated a significant episode-to episode agreement (71.9%) between the two methods, higher in negative samples (89.14%), and a specificity of 75.89%. Clinical variables that gave a statistically significant contribution to their concordance were absolute neutrophil count, ongoing antimicrobial therapy, timing of test execution, and organ localization of infection. The large analysis highlights the potential of molecular-based assays directly performed on blood samples, especially if implementing the detection of antibiotic resistance genes, which was lacking in the used study.

  • Subscribe to Bone Marrow Transplantation for full access:



Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.


  1. 1.

    Gonsalves MD, Sakr Y. Early identification of sepsis. Curr Infect Dis Rep. 2010;12:329–35.

  2. 2.

    Angus DC, van der Poll T. Severe sepsis and septic shock. N Engl J Med. 2013;369:840–51.

  3. 3.

    Gooley TA, Chien JW, Pergam SA, Hingorani S, Sorror ML, Boeckh M, et al. Reduced mortality after allogeneic hematopoietic-cell transplantation. N Engl J Med. 2010;363:2091–101.

  4. 4.

    Morrissey CO, Chen SC, Sorrell TC, Milliken S, Bardy PG, Bradstock KF, et al. Galactomannan and PCR versus culture and histology for directing use of antifungal treatment for invasive aspergillosis in high-risk haematology patients: a randomised controlled trial. Lancet Infect Dis. 2013;13:519–28.

  5. 5.

    Gratwohl A, Brand R, Frassoni F, Rocha V, Niederwieser D, Reusser P, et al. Cause of death after allogeneic haematopoietic stem cell transplantation (HSCT) in early leukaemias: an EBMT analysis of lethal infectious complications and changes over calendar time. Bone Marrow Transplant. 2005;36:757–69.

  6. 6.

    Mancini N, Carletti S, Ghidoli N, Cichero P, Burioni R, Clementi M. The era of molecular and other non-culture-based methods in diagnosis of sepsis. Clin Microbiol Rev. 2010;23:235–51.

  7. 7.

    Wallet F, Nseir S, Baumann L, Herwegh S, Sendid B, Boulo M, et al. Preliminary clinical study using a multiplex real-time PCR test for the detection of bacterial and fungal DNA directly in blood. Clin Microbiol Infect. 2010;16:774–9.

  8. 8.

    Mancini N, Clerici D, Diotti R, Perotti M, Ghidoli N, De Marco D, et al. Molecular diagnosis of sepsis in neutropenic patients with haematological malignancies. J Med Microbiol. 2008;57:601–4.

  9. 9.

    Varani S, Stanzani M, Paolucci M, Melchionda F, Castellani G, Nardi L, et al. Diagnosis of bloodstream infections in immunocompromised patients by real-time PCR. J Infect. 2009;58:346–51.

  10. 10.

    Bravo D, Blanquer J, Tormo M, Aguilar G, Borras R, Solano C, et al. 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. 2011;15:e326–31.

  11. 11.

    Guido M, Quattrocchi M, Zizza A, Pasanisi G, Pavone V, Lobreglio G, et al. Molecular approaches in the diagnosis of sepsis in neutropenic patients with haematological malignances. J Prev Med Hyg. 2012;53:104–8.

  12. 12.

    Reers Y, Idelevich EA, Patkau H, Sauerland MC, Tafelski S, Nachtigall I, et al. Multiplex PCR assay underreports true bloodstream infections with coagulase-negative staphylococci in hematological patients with febrile neutropenia. Diagn Microbiol Infect Dis. 2016;85:413–5.

  13. 13.

    Dark P, Wilson C, Blackwood B, McAuley DF, Perkins GD, McMullan R, et al. 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. 2012;2:e000392.

  14. 14.

    Lamoth F, Jaton K, Prod’hom G, Senn L, Bille J, Calandra T, et al. Multiplex blood PCR in combination with blood cultures for improvement of microbiological documentation of infection in febrile neutropenia. J Clin Microbiol. 2010;48:3510–6.

  15. 15.

    Zaia J, Baden L, Boeckh MJ, Chakrabarti S, Einsele H, Ljungman P, et al. Viral disease prevention after hematopoietic cell transplantation. Bone Marrow Transplant. 2009;44:471–82.

  16. 16.

    Maertens J, Marchetti O, Herbrecht R, Cornely OA, Fluckiger U, Frere P, et al. European guidelines for antifungal management in leukemia and hematopoietic stem cell transplant recipients: summary of the ECIL 3--2009 update. Bone Marrow Transplant. 2011;46:709–18.

  17. 17.

    Freifeld AG, Bow EJ, Sepkowitz KA, Boeckh MJ, Ito JI, Mullen CA, et al. Clinical practice guideline for the use of antimicrobial agents in neutropenic patients with cancer: 2010 update by the infectious diseases society of America. Clin Infect Dis. 2011;52:e56–93.

  18. 18.

    Young JA, Logan BR, Wu J, Wingard JR, Weisdorf DJ, Mudrick C, et al. Infections after transplantation of bone marrow or peripheral blood stem cells from unrelated donors. Biol Blood Marrow Transplant. 2016;22:359–70.

  19. 19.

    De Pauw B, Walsh TJ, Donnelly JP, Stevens DA, Edwards JE, Calandra T, et al. Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group. Clin Infect Dis. 2008;46:1813–21.

  20. 20.

    Wisplinghoff H, Seifert H, Wenzel RP, Edmond MB. Current trends in the epidemiology of nosocomial bloodstream infections in patients with hematological malignancies and solid neoplasms in hospitals in the United States. Clin Infect Dis. 2003;36:1103–10.

  21. 21.

    Ruhnke M, Arnold R, Gastmeier P. Infection control issues in patients with haematological malignancies in the era of multidrug-resistant bacteria. Lancet Oncol. 2014;15:e606–19.

  22. 22.

    Balletto E, Mikulska M. Bacterial infections in hematopoietic stem cell transplant recipients. Mediterr J Hematol Infect Dis. 2015;7:e2015045.

  23. 23.

    Klastersky J, Ameye L, Maertens J, Georgala A, Muanza F, Aoun M, et al. Bacteraemia in febrile neutropenic cancer patients. Int J Antimicrob Agents. 2007;30(Suppl 1):S51–9.

  24. 24.

    Girmenia C, Rossolini GM, Piciocchi A, Bertaina A, Pisapia G, Pastore D, et al. Infections by carbapenem-resistant Klebsiella pneumoniae in SCT recipients: a nationwide retrospective survey from Italy. Bone Marrow Transplant. 2015;50:282–8.

  25. 25.

    Dark P, Blackwood B, Gates S, McAuley D, Perkins GD, McMullan R, et al. Accuracy of LightCycler((R)) 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. 2015;41:21–33.

  26. 26.

    Lehmann LE, Hunfeld KP, Emrich T, Haberhausen G, Wissing H, Hoeft A, et al. A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples. Med Microbiol Immunol. 2008;197:313–24.

  27. 27.

    Mancini N, Sambri V, Corti C, Ghidoli N, Tolomelli G, Paolucci M, et al. Cost-effectiveness of blood culture and a multiplex real-time PCR in hematological patients with suspected sepsis: an observational propensity score-matched study. Expert Rev Mol Diagn. 2014;14:623–32.

  28. 28.

    Lagu T, Rothberg MB, Shieh MS, Pekow PS, Steingrub JS, Lindenauer PK. Hospitalizations, costs, and outcomes of severe sepsis in the United States 2003 to 2007. Crit Care Med. 2012;40:754–61.

  29. 29.

    Greco R, Mancini N, Peccatori J, Cieri N, Vago L, Giglio F, et al. Early molecular diagnosis of aspergillosis in a patient with acute myeloid leukaemia. Heart, lung Vessels. 2014;6:119–24.

Download references


Support was provided by the Italian Ministry of Health, University and Research.

Author contributions

R.G, M.C.B, N.M, M.C, and F.C. conceived the study, analyzed data and wrote the manuscript. All authors carried out the collection of patient information and provided clinical care. All authors read and approved the final manuscript.

Author information

Author notes

  1. Raffaella Greco, Maria Chiara Barbanti, Massimo Clementi and Fabio Ciceri contributed equally to this work.


  1. Hematology and BMT Unit, IRCCS San Raffaele Scientific Institute, via Olgettina 60, 20132, Milano, Italy

    • Raffaella Greco
    • , Maria Chiara Barbanti
    • , Lara Crucitti
    • , Alessandra Forcina
    • , Francesca Lorentino
    • , Luca Vago
    • , Carlo Messina
    • , Daniela Clerici
    • , Mara Morelli
    • , Fabio Giglio
    • , Maria Teresa Lupo Stanghellini
    • , Matteo G. Carrabba
    • , Sarah Marktel
    • , Andrea Assanelli
    • , Massimo Bernardi
    • , Jacopo Peccatori
    • , Consuelo Corti
    •  & Fabio Ciceri
  2. Laboratory of Microbiology and Virology, IRCCS San Raffaele Scientific Institute, Milano, Italy

    • Nicasio Mancini
    • , Laura Infurnari
    •  & Massimo Clementi
  3. Università degli Studi di Milano, Milano, MI, Italy

    • Lara Crucitti
  4. Infectious Disease Unit, IRCCS San Raffaele Scientific Institute, Milano, Italy

    • Chiara Oltolini
  5. Unit of Immunogenetics, Leukemia Genomics and Immunobiology IRCCS San Raffaele Scientific Institute, Milano, Italy

    • Luca Vago
    •  & Paolo Scarpellini
  6. University Vita-Salute, via Olgettina 48, 20132, Milano, Italy

    • Massimo Clementi
    •  & Fabio Ciceri


  1. Search for Raffaella Greco in:

  2. Search for Maria Chiara Barbanti in:

  3. Search for Nicasio Mancini in:

  4. Search for Lara Crucitti in:

  5. Search for Chiara Oltolini in:

  6. Search for Alessandra Forcina in:

  7. Search for Francesca Lorentino in:

  8. Search for Luca Vago in:

  9. Search for Carlo Messina in:

  10. Search for Daniela Clerici in:

  11. Search for Mara Morelli in:

  12. Search for Fabio Giglio in:

  13. Search for Maria Teresa Lupo Stanghellini in:

  14. Search for Laura Infurnari in:

  15. Search for Matteo G. Carrabba in:

  16. Search for Sarah Marktel in:

  17. Search for Andrea Assanelli in:

  18. Search for Paolo Scarpellini in:

  19. Search for Massimo Bernardi in:

  20. Search for Jacopo Peccatori in:

  21. Search for Consuelo Corti in:

  22. Search for Massimo Clementi in:

  23. Search for Fabio Ciceri in:

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Fabio Ciceri.