Rapid extraction of fungal DNA from clinical samples for PCR amplification.
A hexadecyltrimethylammonium bromide (CTAB) method for isolating fungal DNA from clinical samples, suitable for PCR amplification is described. Yeast and filamentous fungi DNA from clinical samples was amplified with primers complementary to the genes coding for rRNA, amplifying a 105 bp fragment and internal transcribed spacer primers amplifying fragments between 242 and 622 bp. The level of sensitivity was 10 +/- 5 yeast and 28 Aspergillus fumigatus CFU ml-1 of biological fluid.
Velegraki
A
A
Department of Microbiology, Medical School, University of Athens, Greece. avelegra@acropolis.gr
Kambouris
M
M
Kostourou
A
A
Chalevelakis
G
G
Legakis
N J
NJ
D001769:Blood / Q000382:microbiology*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D015169:Colony Count, Microbial; D017931:DNA Primers; D004271:DNA, Fungal / Q000302:isolation & purification*; D016469:Fungemia / Q000382:microbiology*; D020459:Genes, rRNA; D006801:Humans; D003904:Mitosporic Fungi / Q000254:growth & development / Q000302:isolation & purification; D009181:Mycoses / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D012680:Sensitivity and Specificity
D002593:Cetrimonium Compounds; D017931:DNA Primers; D004271:DNA, Fungal; D012335:RNA, Ribosomal; D000077286:Cetrimonium