title: Diagnosis by real-time polymerase chain reaction of pathogens and antimicrobial resistance genes in bone marrow transplant patients with bloodstream infections.
abstract: BACKGROUND
  Early identification of pathogens and antimicrobial resistance in bloodstream infections (BSIs) decreases morbidity and mortality, particularly in immunocompromised patients. The aim of the present study was to compare real-time polymerase chain reaction (PCR) with commercial kits for detection of 17 pathogens from blood culture (BC) and 10 antimicrobial resistance genes.
  
  
  METHODS
  A total of 160 BCs were taken from bone marrow transplant patients and screened with Gram-specific probes by multiplex real-time PCR and 17 genus-specific sequences using TaqMan probes and blaSHV, blaTEM, blaCTX, blaKPC, blaIMP, blaSPM, blaVIM, vanA, vanB, and mecA genes by SYBR Green.
  
  
  RESULTS
  Twenty-three of 33 samples identified by phenotypic testing were concordantly positive by BC and real-time PCR. Pathogen identification was discordant in 13 cases. In 12 of 15 coagulase-negative staphylococci, the mecA gene was detected and four Enterococcus spp. were positive for vanA. Two blaCTX and three blaSHV genes were found by quantitative PCR. The blaKPC and metallo-β-lactamase genes were not detected. Five fungal species were identified only by real-time PCR.
  
  
  CONCLUSIONS
  Real-time PCR could be a valuable complementary tool in the management of BSI in bone marrow transplants patients, allowing identification of pathogens and antimicrobial resistance genes.
authors.affiliation: Federal University of São Paulo/UNIFESP, São Paulo, Brazil.
authors.forename: Liana Carballo
authors.initials: LC
authors.lastname: Menezes
authors.forename: Talita Trevizani
authors.initials: TT
authors.lastname: Rocchetti
authors.forename: Karen de Castro
authors.initials: Kde C
authors.lastname: Bauab
authors.forename: Paola
authors.initials: P
authors.lastname: Cappellano
authors.forename: Milene Gonçalves
authors.initials: MG
authors.lastname: Quiles
authors.forename: Fabianne
authors.initials: F
authors.lastname: Carlesse
authors.forename: Jose Salvador Rodrigues
authors.initials: JS
authors.lastname: de Oliveira
authors.forename: Antonio Carlos Campos
authors.initials: AC
authors.lastname: Pignatari
mesh_terms: D000328:Adult; D001419:Bacteria / Q000145:classification / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D016026:Bone Marrow Transplantation / Q000009:adverse effects; D002648:Child; D024881:Drug Resistance, Bacterial; D025141:Drug Resistance, Fungal; D005658:Fungi / Q000145:classification / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016867:Immunocompromised Host; D008828:Microbiological Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018805:Sepsis / Q000382:microbiology*
references.citation: J Clin Microbiol. 2010 Jun;48(6):2030-6
references.pmid: 20392911
references.citation: Crit Care Med. 2008 May;36(5):1487-92
references.pmid: 18434904
references.citation: J Clin Microbiol. 2006 Jun;44(6):1977-81
references.pmid: 16757587
references.citation: J Intern Med. 1998 Nov;244(5):379-86
references.pmid: 9845853
references.citation: Diagn Microbiol Infect Dis. 2004 Sep;50(1):59-69
references.pmid: 15380279
references.citation: J Clin Microbiol. 2010 Oct;48(10):3510-6
references.pmid: 20720024
references.citation: Int J Infect Dis. 2009 Jan;13(1):97-102
references.pmid: 18672389
references.citation: Crit Care Med. 2003 Dec;31(12):2742-51
references.pmid: 14668610
references.citation: J Infect. 2009 May;58(5):346-51
references.pmid: 19362374
references.citation: J Clin Microbiol. 2011 May;49(5):1866-71
references.pmid: 21411591
references.citation: J Clin Microbiol. 2007 Feb;45(2):544-7
references.pmid: 17093019
references.citation: Rev Infect Dis. 1986 Sep-Oct;8(5):792-802
references.pmid: 3538319
references.citation: Med Microbiol Immunol. 2008 Sep;197(3):313-24
references.pmid: 18008085
references.citation: J Clin Microbiol. 2010 Jan;48(1):26-33
references.pmid: 19846634
references.citation: Lancet Infect Dis. 2004 Dec;4(12):751-60
references.pmid: 15567125
references.citation: Br J Haematol. 2000 Nov;111(2):635-40
references.pmid: 11122112
references.citation: J Med Microbiol. 2008 May;57(Pt 5):601-604
references.pmid: 18436593
references.citation: J Clin Microbiol. 2004 Jan;42(1):115-8
references.pmid: 14715740
references.citation: Invest Ophthalmol Vis Sci. 2011 Feb 22;52(2):873-81
references.pmid: 20702819
references.citation: Rinsho Ketsueki. 2010 Jun;51(6):398-401
references.pmid: 20622485
references.citation: Natl Vital Stat Rep. 2008 Apr 24;56(10):1-120
references.pmid: 18512336
references.citation: Clin Chem. 2004 Aug;50(8):1301-14
references.pmid: 15166107
references.citation: Acta Paediatr. 2008 Oct;97(10):1376-80
references.pmid: 18624992
references.citation: J Clin Microbiol. 2008 Aug;46(8):2613-9
references.pmid: 18550744
references.citation: J Med Microbiol. 2006 Jan;55(Pt 1):53-57
references.pmid: 16388030