Use of 65 kDa mannoprotein gene primers in Real Time PCR identification of Candida albicans in biological samples.
A method for the detection and quantification of Candida albicans in biological samples (blood, urine and serum) was developed with the use of Real-Time PCR utilizing CaMP65-specific primers. Two different systems were used for the detection in the LightCycler platform (Roche): the SYBR green fluorescent dye with melting peak analysis and the 5'nuclease fluorescent-probe detection. The amplification was highly specific for C. albicans, providing no cross-reaction on genomic DNA extracted from other Candida species or Aspergillus. The sensitivity in simulated biological samples was especially high (1 genome) when applied to sera and urine, and in blood samples the limit of detection was higher by ten-fold. Finally, the real-time PCR was employed in order to detect and quantify C. albicans in the sera from patients with invasive candidiasis.
Arancia
Silvia
S
Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
Carattoli
Alessandra
A
La Valle
Roberto
R
Cassone
Antonio
A
De Bernardis
Flavia
F
D001230:Aspergillus / Q000235:genetics; D002175:Candida / Q000235:genetics; D002176:Candida albicans / Q000145:classification* / Q000378:metabolism; D002177:Candidiasis / Q000175:diagnosis / Q000382:microbiology; D017931:DNA Primers; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000097:blood / Q000235:genetics* / Q000652:urine; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004271:DNA, Fungal; D008562:Membrane Glycoproteins; C025363:mannoproteins