Detection of Candida albicans DNA with a yeast-specific primer system by polymerase chain reaction.
The in vitro and in vivo selectivity and sensitivity of a yeast-specific primer system was investigated. A two-step polymerase chain reaction (PCR) was used: the first amplified a 245-bp fragment of the gene for cytochrome P450L1A1 and the second a product of 193 bp. This nested PCR produced an approximately 1000-fold increase in the sensitivity of the test for Candida albicans DNA compared with the first primer pair. The lower level of sensitivity of the test in physiological saline and tissue homogenate was about 10 C. albicans cells ml-1. On the other hand, the sensitivity of the nested PCR method was reduced by a factor of more than 1000 when C. albicans was fixed with 4% formalin. After i.v. injection of different doses of C. albicans into mice, the yeast could be demonstrated in blood and in six different organs. The nested PCR was to some extent more sensitive than culturing for the detection of the yeast in the specimens of organs such as lung, cardiac muscle, liver, kidneys and brain. In contrast, in blood and spleen the culture was superior to the PCR technique used. Nested PCR is thus a useful additional method for the demonstration of yeasts.
Wildfeuer
A
A
Research and Development, Pfizer/Mack, Illertissen, Germany.
Schlenk
R
R
Friedrich
W
W
D000818:Animals; D002176:Candida albicans / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis*; D005800:Genes, Fungal; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004271:DNA, Fungal; D003577:Cytochrome P-450 Enzyme System