Clinical Microbiology Newsletter
Volume 32, Issue 6 , Pages 41-49 , 15 March 2010

Code Sepsis: Rapid Methods To Diagnose Sepsis and Detect Hematopathogens: Part II: Challenges to the Laboratory Diagnosis of Sepsis

  • Donna M. Wolk, MHA, Ph.D., D(ABMM)

      Affiliations

    • Corresponding Author InformationMailing address: Donna M. Wolk, MHA, Ph.D., D(ABMM), Assistant Professor, Department of Pathology, University of Arizona, 1501 N. Campbell Ave., P.O. Box 245059, Tucson, AZ 85724-5059. Tel.: 520-626-3676. Fax:520-694-7329
    • ,
  • Albert B. Fiorello, M.D.

References 

  1. Fenollar F , Raoult D . Molecular diagnosis of bloodstream infections caused by non-cultivable bacteria . Int. J. Antimicrob. Agents . 2007;30(Suppl. 1):S7–S15
  2. Kumar A , et al.   Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock . Crit. Care Med. . 2006;34:1589–1596
  3. Diab M , El-Damarawy M , Shemis M . Rapid identification of methicillin-resistant staphylococci bacteremia among intensive care unit patients . Medscape J. Med. . 2008;10:126
  4. Sturm PD , et al.   Performance of two tube coagulase methods for rapid identification of Staphylococcus aureus from blood cultures and their impact on antimicrobial management . Clin. Microbiol. Infect. . 2008;14:510–513
  5. Bennett K , Sharp SE . Rapid differentiation of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus from blood cultures by use of a direct cefoxitin disk diffusion test . J Clin. Microbiol. . 2008;46:3836–3838
  6. Qian Q , Eichelberger K , Kirby JE . Rapid identification of Staphylococcus aureus in blood cultures by use of the direct tube coagulase test . J. Clin. Microbiol. . 2007;45:2267–2269
  7. Marlowe EM , et al.   Practical therapeutic application of the Oxoid PBP2′ latex agglutination test for the rapid identification of methicillin-resistant Staphylococcus aureus in blood cultures . Am. J. Clin. Pathol. . 2002;118:287–291
  8. Kristensen B , Hojbjerg T , Schonheyder HC . Rapid immunodiagnosis of streptococci and enterococci in blood cultures . APMIS . 2001;109:284–288
  9. Terlecka JA , et al.   Rapid differentiation of Candida albicans from non-albicans species by germ tube test directly from BacTAlert blood culture bottles . Mycoses . 2007;50:48–51
  10. Harriau P , Ruffel F , Lardy JB . Use of BioRad plating agar MRSASelect for the daily detection of methicillin resistant staphylococci isolated from samples taken from blood culture bottles . Pathol. Biol. (Paris) . 2006;54:506–509
  11. Chan WW , Lovgren M , Tyrrell GJ . Utility of the pneumoslide test in identification of Streptococcus pneumoniae in blood cultures and its relation to capsular serotype . J. Clin. Microbiol. . 2009;47:1559–1561
  12. Bruins MJ , et al.   Identification and susceptibility testing of Enterobacteriaceae and Pseudomonas aeruginosa by direct inoculation from positive BACTEC blood culture bottles into Vitek 2 . J Clin. Microbiol. . 2004;42:7–11
  13. Chen JR , et al.   Rapid identification and susceptibility testing using the VITEK 2 system using culture fluids from positive BacT/ALERT blood cultures . J. Microbiol. Immunol. Infect. . 2008;41:259–264
  14. Chung JW , et al.   Evaluation of MicroScan and Phoenix system for rapid identification and susceptibility testing using direct inoculation from positive BACTEC blood culture bottles . Korean J. Lab. Med. . 2009;29:25–34
  15. Funke G , Funke-Kissling P . Use of the BD PHOENIX Automated Microbiology System for direct identification and susceptibility testing of gram-negative rods from positive blood cultures in a three-phase trial . J. Clin. Microbiol. . 2004;42:1466–1470
  16. Ling TK , Liu ZK , Cheng AF . Evaluation of the VITEK 2 system for rapid direct identification and susceptibility testing of gram-negative bacilli from positive blood cultures . J. Clin. Microbiol. . 2003;41:4705–4707
  17. Wellinghausen NT , et al.   Evaluation of the Merlin MICRONAUT system for rapid direct susceptibility testing of gram-positive cocci and gram-negative bacilli from positive blood cultures . J. Clin. Microbiol. . 2007;45:789–795
  18. Avolio M , et al.   Direct antifungal susceptibility testing of positive Candida blood cultures by Sensititre YeastOne . New Microbiol. . 2009;32:179–184
  19. Hunfeld KP , et al.   Molecular biological detection of pathogens in patients with sepsis . Potentials, limitations and perspectives. Anaesthesist . 2008;57:326–337
  20. Osborn TM , Nguyen HB , Rivers EP . Emergency medicine and the surviving sepsis campaign: an international approach to managing severe sepsis and septic shock . Ann. Emerg. Med. . 2005;46:228–231
  21. Rivers E , et al.   Early goal-directed therapy in the treatment of severe sepsis and septic shock . N. Engl. J. Med. . 2001;345:1368–1377
  22. Rivers EP , et al.   Early and innovative interventions for severe sepsis and septic shock: taking advantage of a window of opportunity . Can. Med. Assoc. J. . 2005;173:1054–1065
  23. Adelson ME , et al.   Simultaneous detection of herpes simplex virus types 1 and 2 by real-time PCR and pyrosequencing . J. Clin. Virol. . 2005;33:25–34
  24. Gonzalez V , et al.   Rapid diagnosis of Staphylococcus aureus bacteremia using S. aureus PNA FISH . Eur. J. Clin. Microbiol. Infect. Dis. . 2004;23:396–398
  25. Oliveira K , et al.   Direct identification of Staphylococcus aureus from positive blood culture bottles . J. Clin. Microbiol. . 2003;41:889–891
  26. Oliveira K , et al.   Differentiation of Candida albicans and Candida dubliniensis by fluorescent in situ hybridization with peptide nucleic acid probes . J. Clin. Microbiol. . 2001;39:4138–4141
  27. Oliveira K , et al.   Rapid identification of Staphylococcus aureus directly from blood cultures by fluorescence in situ hybridization with peptide nucleic acid probes . J. Clin. Microbiol. . 2002;40:247–251
  28. Rigby S , et al.   Fluorescence in situ hybridization with peptide nucleic acid probes for rapid identification of Candida albicans directly from blood culture bottles . J. Clin. Microbiol. . 2002;40:2182–2186
  29. Sogaard M , Stender H , Schonheyder HC . Direct identification of major blood culture pathogens, including Pseudomonas aeruginosa and Escherichia coli, by a panel of fluorescence in situ hybridization assays using peptide nucleic acid probes . J. Clin. Microbiol. . 2005;43:1947–1949
  30. Stender H . PNA FISH: an intelligent stain for rapid diagnosis of infectious diseases . Exp. Rev. Mol. Diagn. . 2003;3:649–655
  31. Wilson DA , et al.   Multicenter evaluation of a Candida albicans peptide nucleic acid fluorescent in situ hybridization probe for characterization of yeast isolates from blood cultures . J. Clin. Microbiol. . 2005;43:2909–2912
  32. Alexander BD , et al.   Cost savings with implementation of PNA FISH testing for identification of Candida albicans in blood cultures . Diagn. Microbiol. Infect. Dis. . 2006;54:277–282
  33. Hartmann H , et al.   Rapid identification of Staphylococcus aureus in blood cultures by a combination of fluorescence in situ hybridization using peptide nucleic acid probes and flow cytometry . J. Clin. Microbiol. . 2005;43:4855–4857
  34. Chapin KC , Musgnug MC . Direct susceptibility testing of positive blood cultures by using Sensititre broth microdilution plates . J. Clin. Microbiol. . 2003;41:4751–4754
  35. Oliveria, K.M. et al. 2007. Rapid identification of S. aureus and differentiation between MRSA and MSSA from blood culture bottles using S. aureus PNA FISH and MRSA EVIGENE. Presented at the IDSA 2005 Annual Meeting.
  36. Shepard JR , et al.   Multicenter evaluation of the Candida albicans/Candida glabrata peptide nucleic acid fluorescent in situ hybridization method for simultaneous dual-color identification of C. albicans and C. glabrata directly from blood culture bottles . J. Clin. Microbiol. . 2008;46:50–55
  37. Reller ME , et al.   Use of peptide nucleic acid-fluorescence in situ hybridization for definitive, rapid identification of five common Candida species . J. Clin. Microbiol. . 2007;45:3802–3803
  38. Toombs, L. et al. 2006. Impact of peptide nucleic acid (PNA) fluorescence in situ hybridization (FISH) for enterococcocal blood stream infections. Presented at the IDSA 2006 Annual Meeting.
  39. Forrest GN , et al.   Peptide nucleic acid fluorescent in situ hybridization for hospital-acquired enterococcal bacteremia: delivering earlier effective antimicrobial therapy . Antimicrob. Agents Chemother. . 2008;52:3558–3563
  40. Forrest GN , et al.   Impact of rapid in situ hybridization testing on coagulase-negative staphylococci positive blood cultures . J. Antimicrob. Chemother. . 2006;58:154–158
  41. Ly T , et al.   Impact upon clinical outcomes of translation of PNA FISH-generated laboratory data from the clinical microbiology bench to bedside in real time . Ther. Clin. Risk Manag. . 2008;4:637–640
  42. Pappas PG , et al.   Guidelines for treatment of candidiasis . Clin. Infect. Dis. . 2004;38:161–189
  43. Forrest GN , et al.   Peptide nucleic acid fluorescence in situ hybridization-based identification of Candida albicans and its impact on mortality and antifungal therapy costs . J. Clin. Microbiol. . 2006;44:3381–3383
  44. Wolk DM , et al.   Rapid detection of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in wound specimens and blood cultures: multicenter preclinical evaluation of the Cepheid Xpert MRSA/SA skin and soft tissue and blood culture assays . J. Clin. Microbiol. . 2009;47:823–826
  45. Ito T , et al.   Community-associated methicillin-resistant Staphylococcus aureus: current status and molecular epidemiological perspective . Kansenshogaku Zasshi . 2004;78:459–469
  46. Katayama Y , Zhang HZ , Chambers HF . PBP 2a mutations producing very high-level resistance to beta-lactams . Antimicrob. Agents Chemother. . 2004;48:453–459
  47. Donnio PY , et al.   Partial excision of the chromosomal cassette containing the methicillin resistance determinant results in methicillin-susceptible Staphy lococcus aureus . J. Clin. Microbiol. . 2005;43:4191–4193
  48. Francois P , et al.   Evaluation of three molecular assays for rapid identification of methicillin-resistant Staphylococcus aureus . J. Clin. Microbiol. . 2007;45:2011–2013
  49. Stamper PD , et al.   Clinical validation of the molecular BD GeneOhm StaphSR assay for direct detection of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus in positive blood cultures . J. Clin. Microbiol. . 2007;45:2191–2196
  50. Grobner S , et al.   Evaluation of the BD GeneOhm StaphSR assay for detection of methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates from spiked positive blood culture bottles . J. Clin. Microbiol. . 2009;47:1689–1694
  51. al-Nawas B , Shah PM . Procalcitonin in patients with and without immunosuppression and sepsis . Infection . 1996;24:434–436
  52. Chawes BL , et al.   Procalcitonin for early diagnosis of bacteraemia in children with cancer . Ugeskr. Laeger . 2007;169:138–142
  53. Giamarellos-Bourboulis EJ , et al.   Procalcitonin: a marker to clearly differentiate systemic inflammatory response syndrome and sepsis in the critically ill patient? . Intensive Care Med. . 2002;28:1351–1356
  54. Reingardiene D . Procalcitonin as a marker of the systemic inflammatory response to infection . Medicina . 2004;40:696–701
  55. Sauer M , et al.   Procalcitonin in comparison to C-reactive protein as markers of the course of sepsis in severely immunocompromised children after bone marrow transplantation . Klin. Padiatr. . 2000;212:10–15
  56. Schneider HG , Lam QT . Procalcitonin for the clinical laboratory: a review . Pathology . 2007;39:383–390
  57. Fernandez-Lopez LA , et al.   Procalcitonin in pediatric emergency departments for the early diagnosis of invasive bacterial infections in febrile infants: results of a multicenter study and utility of a rapid qualitative test for this marker . Pediatr. Infect. Dis. J. . 2003;22:895–903
  58. van Rossum AM , Wulkan RW , Oudesluys-Murphy AM . Procalcitonin as an early marker of infection in neonates and children . Lancet Infect. Dis. . 2004;4:620–630
  59. Muller B , et al.   Calcitonin precursors are reliable markers of sepsis in a medical intensive care unit . Crit. Care Med. . 2000;28:977–983
  60. Kruger S , et al.   Procalcitonin predicts patients at low riskof death from community-acquired pneumonia across all CRB-65 classes . Eur. Respir. J. . 2008;31:349–355
  61. Beghetti M , et al.   Kinetics of procalcitonin, interleukin 6 and C-reactive protein after cardiopulmonary-bypass in children . Cardiol. Young. . 2003;13:161–167
  62. Meisner M , Adina H , Schmidt J . Correlation of procalcitonin and C-reactive protein to inflammation, complications, and outcome during the intensive care unit course of multiple-trauma patients . Crit. Care . 2006;10:R1
  63. Brahms Aktiengesellschaft. 2006. Instruction manual BRAHMS PCT sensitive KRYPTOR product insert.
  64. Fenollar F , Raoult D . Molecular diagnosis of bloodstream infections caused by non-cultivable bacteria . Int. J. Antimicrob. Agents . 2007;30(Suppl. 1):S7–S15
  65. Karahan ZC , et al.   PCR evaluation of false-positive signals from two automated blood-culture systems . J. Med. Microbiol. . 2006;55:53–57
  66. Marlowe EM , Wolk DM . Pathogen detection in the genomic era . In:  Tang Y editors. Advanced techniques in diagnostic microbiology . New York: Springer Verlag; 2006;p. 505–523
  67. Wolk DM , Marlowe EM . Future trends in diagnosis of infections in the immunocompromised population . In:  Hayden RT , et al. editor. Diagnostic microbiology of the immunocompromised Host . Washington, DC: ASM Press; 2008;
  68. Casalta JP , et al.   Evaluation of the LightCycler SeptiFast test in the rapid etiologic diagnostic of infectious endocarditis . Eur. J. Clin. Microbiol. Infect. Dis. . 2009;28:569–573
  69. Dierkes C , et al.   Clinical impact of a commercially available multiplex PCR system for rapid detection of pathogens in patients with presumed sepsis . BMC Infect. Dis. . 2009;9:126
  70. Mancini N , et al.   Molecular diagnosis of sepsis in neutropenic patients with haematological malignancies . J. Med. Microbiol. . 2008;57:601–604
  71. Jordan JA , Butchko AR , Durso MB . Use of pyrosequencing of 16S rRNA fragments to differentiate between bacteria responsible for neonatal sepsis . J. Mol. Diagn. . 2005;7:105–110
  72. Tang YW , et al.   StaphPlex system for rapid and simultaneous identification of antibiotic resistance determinants and Panton-Valentine leukocidin detection of staphylococci from positive blood cultures . J. Clin. Microbiol. . 2007;45:1867–1873
  73. Baldwin CD , et al.   Usefulness of multilocus polymerase chain reaction followed by electrospray ionization mass spectrometry to identify a diverse panel of bacterial isolates . Diagn. Microbiol. Infect. Dis. . 2009;63:403–408
  74. Ecker DJ , et al.   Molecular genotyping of microbes by multilocus PCR and mass spectrometry: a new tool for hospital infection control and public health surveillance . Methods Mol. Biol. . 2009;551:71–87
  75. Ecker DJ , et al.   Ibis T5000: a universal biosensor approach for microbiology . Nat. Rev. Microbiol. . 2008;6:553–558
  76. Hujer KM , et al.   Rapid determination of quinolone resistance in Acinetobacter spp . J. Clin. Microbiol. . 2009;47:1436–1442
  77. Hall TA , et al.   Rapid molecular genotyping and clonal complex assignment of Staphylococcus aureus isolates by PCR coupled to electrospray ionization-mass spectrometry . J. Clin. Microbiol. . 2009;47:1733–1741
  78. Wolk DM , et al.   Rapid molecular characterization of S. aureus isolates by PCR/ESI-MS and correlation with phenotype (accpeted for publication) . J. Clin. Microbiol. . 2009;
  79. Diekema DJ , et al.   Rapid detection of antimicrobial-resistant organism carriage: an unmet clinical need . J. Clin. Microbiol. . 2004;42:2879–2883
  80. Fluit AC , Visser MR , Schmitz FJ . Molecular detection of antimicrobial resistance . Clin. Microbiol. Rev. . 2001;14:836–871
  81. Rasheed JK , Tenover FC . Detection and characterization of antimicrobial resistance genes in bacteria . In:  Murray PR , et al. editor. Manual of clinical microbiology . Washington, DC: ASM Press; 2003;p. 1196–1213
  82. Tenover FC , Rasheed JK . Detection of antimicrobial resistance genes and mutations associated with antimicrobial resistance in microorganisms . In:  Persing DH , et al. editor. Molecular microbiology: diagnostic principles and practice . Washington, DC: ASM Press; 2004;p. 391–406
  83. Edlind TD . Molecular detection of antifungal resistance . In:  Persing DH , et al. editor. Molecular microbiology: diagnostic principles and practice . Washington, DC: ASM Press; 2004;p. 569–575

 Editor's Note: Part I of this article appeared in the March 1, 2010 issue of CMN, Volume 32, Number 5.

PII: S0196-4399(10)00010-3

doi: 10.1016/j.clinmicnews.2010.03.001

Clinical Microbiology Newsletter
Volume 32, Issue 6 , Pages 41-49 , 15 March 2010

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