#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Multiplex Identification of Gram-Positive Bacteria and Resistance Determinants Directly from Positive Blood Culture Broths: Evaluation of an Automated Microarray-Based Nucleic Acid Test


Background:
A multicenter study was conducted to evaluate the diagnostic accuracy (sensitivity and specificity) of the Verigene Gram-Positive Blood Culture Test (BC-GP) test to identify 12 Gram-positive bacterial gene targets and three genetic resistance determinants directly from positive blood culture broths containing Gram-positive bacteria.

Methods and Findings:
1,252 blood cultures containing Gram-positive bacteria were prospectively collected and tested at five clinical centers between April, 2011 and January, 2012. An additional 387 contrived blood cultures containing uncommon targets (e.g., Listeria spp., S. lugdunensis, vanB-positive Enterococci) were included to fully evaluate the performance of the BC-GP test. Sensitivity and specificity for the 12 specific genus or species targets identified by the BC-GP test ranged from 92.6%–100% and 95.4%–100%, respectively. Identification of the mecA gene in 599 cultures containing S. aureus or S. epidermidis was 98.6% sensitive and 94.3% specific compared to cefoxitin disk method. Identification of the vanA gene in 81 cultures containing Enterococcus faecium or E. faecalis was 100% sensitive and specific. Approximately 7.5% (87/1,157) of single-organism cultures contained Gram-positive bacteria not present on the BC-GP test panel. In 95 cultures containing multiple organisms the BC-GP test was in 71.6% (68/95) agreement with culture results. Retrospective analysis of 107 separate blood cultures demonstrated that identification of methicillin resistant S. aureus and vancomycin resistant Enterococcus spp. was completed an average of 41.8 to 42.4 h earlier using the BC-GP test compared to routine culture methods. The BC-GP test was unable to assign mecA to a specific organism in cultures containing more than one Staphylococcus isolate and does not identify common blood culture contaminants such as Micrococcus, Corynebacterium, and Bacillus.

Conclusions:
The BC-GP test is a multiplex test capable of detecting most leading causes of Gram-positive bacterial blood stream infections as well as genetic markers of methicillin and vancomycin resistance directly from positive blood cultures.

Please see later in the article for the Editors' Summary


Vyšlo v časopise: Multiplex Identification of Gram-Positive Bacteria and Resistance Determinants Directly from Positive Blood Culture Broths: Evaluation of an Automated Microarray-Based Nucleic Acid Test. PLoS Med 10(7): e32767. doi:10.1371/journal.pmed.1001478
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pmed.1001478

Souhrn

Background:
A multicenter study was conducted to evaluate the diagnostic accuracy (sensitivity and specificity) of the Verigene Gram-Positive Blood Culture Test (BC-GP) test to identify 12 Gram-positive bacterial gene targets and three genetic resistance determinants directly from positive blood culture broths containing Gram-positive bacteria.

Methods and Findings:
1,252 blood cultures containing Gram-positive bacteria were prospectively collected and tested at five clinical centers between April, 2011 and January, 2012. An additional 387 contrived blood cultures containing uncommon targets (e.g., Listeria spp., S. lugdunensis, vanB-positive Enterococci) were included to fully evaluate the performance of the BC-GP test. Sensitivity and specificity for the 12 specific genus or species targets identified by the BC-GP test ranged from 92.6%–100% and 95.4%–100%, respectively. Identification of the mecA gene in 599 cultures containing S. aureus or S. epidermidis was 98.6% sensitive and 94.3% specific compared to cefoxitin disk method. Identification of the vanA gene in 81 cultures containing Enterococcus faecium or E. faecalis was 100% sensitive and specific. Approximately 7.5% (87/1,157) of single-organism cultures contained Gram-positive bacteria not present on the BC-GP test panel. In 95 cultures containing multiple organisms the BC-GP test was in 71.6% (68/95) agreement with culture results. Retrospective analysis of 107 separate blood cultures demonstrated that identification of methicillin resistant S. aureus and vancomycin resistant Enterococcus spp. was completed an average of 41.8 to 42.4 h earlier using the BC-GP test compared to routine culture methods. The BC-GP test was unable to assign mecA to a specific organism in cultures containing more than one Staphylococcus isolate and does not identify common blood culture contaminants such as Micrococcus, Corynebacterium, and Bacillus.

Conclusions:
The BC-GP test is a multiplex test capable of detecting most leading causes of Gram-positive bacterial blood stream infections as well as genetic markers of methicillin and vancomycin resistance directly from positive blood cultures.

Please see later in the article for the Editors' Summary


Zdroje

1. AngusDC, WaxRS (2001) Epidemiology of sepsis: an update. Crit Care Med 29: S109–S116.

2. MartinGS, ManninoDM, EatonS, MossM (2003) The epidemiology of sepsis in the United States from 1979 through 2000. N Engl J Med 348: 1546–1554.

3. LevyMM, ArtigasA, PhillipsGS, RhodesA, BealeR, et al. (2012) Outcomes of the Surviving Sepsis Campaign in intensive care units in the USA and Europe: a prospective cohort study. Lancet Infect Dis 12: 919–924.

4. PittetD, TararaD, WenzelRP (1994) Nosocomial bloodstream infection in critically ill patients. Excess length of stay, extra costs, and attributable mortality. JAMA 271: 1598–1601.

5. GrozdanovskiK, MilenkovicZ, DemiriI, SpasovskaK (2012) Prediction of outcome from community-acquired severe sepsis and septic shock in tertiary-care university hospital in a developing country. Crit Care Res Pract 2012: 182324.

6. WisplinghoffH, BischoffT, TallentSM, SeifertH, WenzelRP, et al. (2004) Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Clin Infect Dis 39: 309–317.

7. StonePW, BracciaD, LarsonE (2005) Systematic review of economic analyses of health care-associated infections. Am J Infect Control 33: 501–509.

8. KumarA, RobertsD, WoodKE, LightB, ParrilloJE, et al. (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.

9. BryanCS, ReynoldsKL, BrennerER (1983) Analysis of 1,186 episodes of gram-negative bacteremia in non-university hospitals: the effects of antimicrobial therapy. Rev Infect Dis 5: 629–638.

10. BauerKA, WestJE, Balada-LlasatJM, PancholiP, StevensonKB, et al. (2010) An antimicrobial stewardship program's impact with rapid polymerase chain reaction methicillin-resistant Staphylococcus aureus/S. aureus blood culture test in patients with S. aureus bacteremia. Clin Infect Dis 51: 1074–1080.

11. DellingerRP, LevyMM, CarletJM, BionJ, ParkerMM, et al. (2008) Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock: 2008. Intensive Care Med 34: 17–60.

12. BuchanBW, RiebeKM, LedeboerNA (2012) Comparison of the MALDI Biotyper system using Sepsityper specimen processing to routine microbiological methods for identification of bacteria from positive blood culture bottles. J Clin Microbiol 50: 346–352.

13. La ScolaB, RaoultD (2009) Direct identification of bacteria in positive blood culture bottles by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry. PLoS One 4: e8041 doi:10.1371/journal.pone.0008041

14. StevensonLG, DrakeSK, MurrayPR (2010) Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 48: 444–447.

15. DeckMK, AndersonES, BucknerRJ, ColasanteG, CoullJM, et al. (2012) Multicenter evaluation of the Staphylococcus QuickFISH method for simultaneous identification of Staphylococcus aureus and coagulase-negative staphylococci directly from blood culture bottles in less than 30 minutes. J Clin Microbiol 50: 1994–1998.

16. HensleyDM, TapiaR, EncinaY (2009) An evaluation of the advandx Staphylococcus aureus/CNS PNA FISH assay. Clin Lab Sci 22: 30–33.

17. CattoirV, GilibertA, Le GlaunecJM, LaunayN, Bait-MerabetL, et al. (2010) Rapid detection of Pseudomonas aeruginosa from positive blood cultures by quantitative PCR. Ann Clin Microbiol Antimicrob 9: 21.

18. JukesL, MikhailJ, Bome-MannathokoN, HadfieldSJ, HarrisLG, et al. (2010) Rapid differentiation of Staphylococcus aureus, Staphylococcus epidermidis and other coagulase-negative staphylococci and meticillin susceptibility testing directly from growth-positive blood cultures by multiplex real-time PCR. J Med Microbiol 59: 1456–1461.

19. WolkDM, StruelensMJ, PancholiP, DavisT, Della-LattaP, et al. (2009) 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 47: 823–826.

20. IkryannikovaLN, FilimonovaAV, MalakhovaMV, SavinovaT, FilimonovaO, et al. (2012) Discrimination between Streptococcus pneumoniae and Streptococcus mitis based on sorting of their MALDI mass spectra. Clin Microbiol Infect In press.

21. MartinyD, BussonL, WyboI, El HajRA, DedisteA, et al. (2012) Comparison of the Microflex LT and Vitek MS systems for routine identification of bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 50: 1313–1325.

22. WernoAM, ChristnerM, AndersonTP, MurdochDR (2012) Differentiation of Streptococcus pneumoniae from nonpneumococcal streptococci of the Streptococcus mitis group by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 50: 2863–2867.

23. ElghanianR, StorhoffJJ, MucicRC, LetsingerRL, MirkinCA (1997) Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. Science 277: 1078–1081.

24. StorhoffJJ, MarlaSS, BaoP, HagenowS, MehtaH, et al. (2004) Gold nanoparticle-based detection of genomic DNA targets on microarrays using a novel optical detection system. Biosens Bioelectron 19: 875–883.

25. CLSI (2011) Performance standards for antimicrobial susceptibility testing; 21st informational supplement. M100-S21.

26. PauleSM, PasquarielloAC, ThomsonRBJr, KaulKL, PetersonLR (2005) Real-time PCR can rapidly detect methicillin-susceptible and methicillin-resistant Staphylococcus aureus directly from positive blood culture bottles. Am J Clin Pathol 124: 404–407.

27. ClarridgeJE3rd (2004) Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev 17: 840–862 table of contents.

28. KhamisA, RaoultD, La ScolaB (2005) Comparison between rpoB and 16S rRNA gene sequencing for molecular identification of 168 clinical isolates of Corynebacterium. J Clin Microbiol 43: 1934–1936.

29. TamuraK, PetersonD, PetersonN, StecherG, NeiM, et al. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731–2739.

30. PetersRP, SavelkoulPH, Simoons-SmitAM, DannerSA, Vandenbroucke-GraulsCM, et al. (2006) Faster identification of pathogens in positive blood cultures by fluorescence in situ hybridization in routine practice. J Clin Microbiol 44: 119–123.

31. MorganMA, MarloweE, Novak-WeeklyS, MillerJM, PainterTM, et al. (2011) A 1.5 hour procedure for identification of Enterococcus Species directly from blood cultures. J Vis Exp pii: 2616.

32. ShepardJR, AddisonRM, AlexanderBD, Della-LattaP, GhernaM, et al. (2008) 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 46: 50–55.

33. StevensonLG, DrakeSK, SheaYR, ZelaznyAM, MurrayPR (2010) Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of clinically important yeast species. J Clin Microbiol 48: 3482–3486.

34. BurckhardtI, ZimmermannS (2011) Using matrix-assisted laser desorption ionization-time of flight mass spectrometry to detect carbapenem resistance within 1 to 2.5 hours. J Clin Microbiol 49: 3321–3324.

35. DuZ, YangR, GuoZ, SongY, WangJ (2002) Identification of Staphylococcus aureus and determination of its methicillin resistance by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Anal Chem 74: 5487–5491.

36. TissariP, ZumlaA, TarkkaE, MeroS, SavolainenL, et al. (2010) Accurate and rapid identification of bacterial species from positive blood cultures with a DNA-based microarray platform: an observational study. Lancet 375: 224–230.

37. HallKK, LymanJA (2006) Updated review of blood culture contamination. Clin Microbiol Rev 19: 788–802.

38. IbrahimEH, ShermanG, WardS, FraserVJ, KollefMH (2000) The influence of inadequate antimicrobial treatment of bloodstream infections on patient outcomes in the ICU setting. Chest 118: 146–155.

39. RuimyR, Dos-SantosM, RaskineL, BertF, MassonR, et al. (2008) Accuracy and potential usefulness of triplex real-time PCR for improving antibiotic treatment of patients with blood cultures showing clustered gram-positive cocci on direct smears. J Clin Microbiol 46: 2045–2051.

40. PartaM, GoebelM, ThomasJ, MatloobiM, StagerC, et al. (2010) Impact of an assay that enables rapid determination of Staphylococcus species and their drug susceptibility on the treatment of patients with positive blood culture results. Infect Control Hosp Epidemiol 31: 1043–1048.

41. BartelsMD, BoyeK, RohdeSM, LarsenAR, TorfsH, et al. (2009) A common variant of staphylococcal cassette chromosome mec type IVa in isolates from Copenhagen, Denmark, is not detected by the BD GeneOhm methicillin-resistant Staphylococcus aureus assay. J Clin Microbiol 47: 1524–1527.

42. SnyderJW, MunierGK, HeckmanSA, CampP, OvermanTL (2009) Failure of the BD GeneOhm StaphSR assay for direct detection of methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates in positive blood cultures collected in the United States. J Clin Microbiol 47: 3747–3748.

43. KlotchkoA, WallaceMR, LicitraC, SiegerB (2011) Staphylococcus lugdunensis: an emerging pathogen. South Med J 104: 509–514.

44. FrankKL, Del PozoJL, PatelR (2008) From clinical microbiology to infection pathogenesis: how daring to be different works for Staphylococcus lugdunensis. Clin Microbiol Rev 21: 111–133.

45. MandellLA, WunderinkRG, AnzuetoA, BartlettJG, CampbellGD, et al. (2007) Infectious Diseases Society of America/American Thoracic Society consensus guidelines on the management of community-acquired pneumonia in adults. Clin Infect Dis 44 (Suppl 2) S27–S72.

46. ShahSS, DuganMH, BellLM, GrundmeierRW, FlorinTA, et al. (2011) Blood cultures in the emergency department evaluation of childhood pneumonia. Pediatr Infect Dis J 30: 475–479.

47. AbeT, TokudaY, IshimatsuS, BirrerRB (2009) Usefulness of initial blood cultures in patients admitted with pneumonia from an emergency department in Japan. J Infect Chemother 15: 180–186.

48. ChaiwarithR, JullaketW, BunchooM, NuntachitN, SirisanthanaT, et al. (2011) Streptococcus agalactiae in adults at Chiang Mai University Hospital: a retrospective study. BMC Infect Dis 11: 149.

49. EdwardsMS, BakerCJ (2005) Group B streptococcal infections in elderly adults. Clin Infect Dis 41: 839–847.

50. DomingoP, BarquetN, AlvarezM, CollP, NavaJ, et al. (1997) Group B streptococcal meningitis in adults: report of twelve cases and review. Clin Infect Dis 25: 1180–1187.

51. ProtonotariouE, DimitrouliaE, PournarasS, PitirigaV, SofianouD, et al. (2010) Trends in antimicrobial resistance of clinical isolates of Enterococcus faecalis and Enterococcus faecium in Greece between 2002 and 2007. J Hosp Infect 75: 225–227.

52. LyT, GuliaJ, PyrgosV, WagaM, ShohamS (2008) 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 4: 637–640.

53. KimM, HeoSR, ChoiSH, KwonH, ParkJS, et al. (2008) Comparison of the MicroScan, VITEK 2, and Crystal GP with 16S rRNA sequencing and MicroSeq 500 v2.0 analysis for coagulase-negative Staphylococci. BMC Microbiol 8: 233.

54. LayerF, GhebremedhinB, ModerKA, KonigW, KonigB (2006) Comparative study using various methods for identification of Staphylococcus species in clinical specimens. J Clin Microbiol 44: 2824–2830.

55. KobayashiN, TaniguchiK, UrasawaS (1998) Analysis of diversity of mutations in the mecI gene and mecA promoter/operator region of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother 42: 717–720.

56. OliveiraDC, de LencastreH (2011) Methicillin-resistance in Staphylococcus aureus is not affected by the overexpression in trans of the mecA gene repressor: a surprising observation. PLoS One 6: e23287 doi:10.1371/journal.pone.0023287

57. BargN, ChambersH, KernodleD (1991) Borderline susceptibility to antistaphylococcal penicillins is not conferred exclusively by the hyperproduction of beta-lactamase. Antimicrob Agents Chemother 35: 1975–1979.

58. MassiddaO, MingoiaM, FaddaD, WhalenMB, MontanariMP, et al. (2006) Analysis of the beta-lactamase plasmid of borderline methicillin-susceptible Staphylococcus aureus: focus on bla complex genes and cadmium resistance determinants cadD and cadX. Plasmid 55: 114–127.

Štítky
Interné lekárstvo

Článok vyšiel v časopise

PLOS Medicine


2013 Číslo 7
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#