title: Rapid identification of yeasts from positive blood culture bottles by pyrosequencing.
abstract: We have developed a rapid protocol for the identification of Candida species from positive blood cultures by combining a simple method for nucleic acid extraction and preparation using microbial storage cardboards with polymerase chain reaction (PCR) and pyrosequencing of a small region of the 18 S rRNA gene. The protocol is robust and easy to implement and can be performed in 4 h. The method was tested against a collection of clinical blood cultures. Agreement of sequence identifications with standard microbiological methods was 100%.
authors.affiliation: Servicio de Microbiología, Hospital Universitario La Paz-IdiPAZ, Paseo de La Castellana, Madrid, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Quiles-Melero
authors.forename: J
authors.initials: J
authors.lastname: García-Rodriguez
authors.forename: M P
authors.initials: MP
authors.lastname: Romero-Gómez
authors.forename: P
authors.initials: P
authors.lastname: Gómez-Sánchez
authors.forename: J
authors.initials: J
authors.lastname: Mingorance
mesh_terms: D001769:Blood / Q000382:microbiology*; D002175:Candida / Q000145:classification* / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis* / Q000382:microbiology; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D016469:Fungemia / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D009172:Mycology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012331:RNA, Fungal / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012331:RNA, Fungal; D012337:RNA, Ribosomal, 18S
references.citation: J Clin Microbiol. 2005 Oct;43(10):5256-62
references.pmid: 16207992
references.citation: Am J Ther. 2009 Nov-Dec;16(6):508-11
references.pmid: 19531934
references.citation: Curr Med Res Opin. 2009 Jul;25(7):1732-40
references.pmid: 19519284
references.citation: Mycoses. 2009 May;52(3):197-205
references.pmid: 19391253
references.citation: Nucleic Acids Res. 2007;35(21):7188-96
references.pmid: 17947321
references.citation: Epidemiol Infect. 2010 Sep;138(9):1328-35
references.pmid: 20056017
references.citation: J Clin Microbiol. 2003 Dec;41(12):5660-4
references.pmid: 14662958
references.citation: J Clin Microbiol. 2002 Aug;40(8):2860-5
references.pmid: 12149343
references.citation: J Antimicrob Chemother. 2009 Sep;64(3):625-9
references.pmid: 19622536
references.citation: Arch Pathol Lab Med. 2008 Apr;132(4):667-74
references.pmid: 18384218
references.citation: Eur J Clin Microbiol Infect Dis. 2003 Nov;22(11):693-6
references.pmid: 14557922
references.citation: Clin Infect Dis. 2009 Feb 1;48(3):265-73
references.pmid: 19115967
references.citation: Eur J Clin Microbiol Infect Dis. 2008 Sep;27(9):821-30
references.pmid: 18421488
references.citation: Clin Infect Dis. 2007 Jun 15;44(12):e109-14
references.pmid: 17516389
references.citation: Clin Infect Dis. 2009 Jun 15;48(12):1695-703
references.pmid: 19441981
references.citation: J Clin Microbiol. 2009 Jan;47(1):247-8
references.pmid: 18987173
references.citation: Mycoses. 2004 Feb;47(1-2):29-33
references.pmid: 14998396