Rapid identification of yeasts from positive blood culture bottles by pyrosequencing.
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%.
Quiles-Melero
I
I
Servicio de Microbiología, Hospital Universitario La Paz-IdiPAZ, Paseo de La Castellana, Madrid, Spain.
García-Rodriguez
J
J
Romero-Gómez
M P
MP
Gómez-Sánchez
P
P
Mingorance
J
J
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
D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012331:RNA, Fungal; D012337:RNA, Ribosomal, 18S
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