title: Improved detection of Candida albicans by PCR in blood of neutropenic mice with systemic candidiasis.
abstract: A PCR using primers aimed at the multicopy gene coding for the small subunit rRNA and resulting in the synthesis of a 180-bp fragment was evaluated for its use in diagnosing invasive candidiasis in comparison with blood culture. With the use of a C. albicans-specific probe, +/- 10 to 15 C. albicans cells are detected in 100 microliters of whole blood by Southern analysis. A DNase pretreatment was critical in the purification process of yeast DNA from whole blood. Omission of the DNase pretreatment decreased assay sensitivity 10-fold. PCR analysis of blood specimens collected from mice with invasive candidiasis is more sensitive than blood culture (100 versus 67%, respectively) at 72 h after intravenous (i.v.) inoculation with C. albicans. Furthermore, the intensity of the hybridization signals increased with the progression of infection. In contrast, multiple blood samples from gastrointestinally colonized mice were all negative by PCR, indicating that the PCR assay is also specific and may, therefore, make a positive contribution to the detection and follow-up of invasive candidiasis.
authors.affiliation: Department of Clinical Microbiology and Antimicrobial Therapy, Erasmus University School of Medicine, Rotterdam, The Netherlands.
authors.forename: A J
authors.initials: AJ
authors.lastname: van Deventer
authors.forename: W H
authors.initials: WH
authors.lastname: Goessens
authors.forename: A
authors.initials: A
authors.lastname: van Belkum
authors.forename: H J
authors.initials: HJ
authors.lastname: van Vliet
authors.forename: E W
authors.initials: EW
authors.lastname: van Etten
authors.forename: H A
authors.initials: HA
authors.lastname: Verbrugh
mesh_terms: D000818:Animals; D001483:Base Sequence; D001769:Blood / Q000382:microbiology; D015139:Blotting, Southern; D002176:Candida albicans / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002177:Candidiasis / Q000382:microbiology*; D015169:Colony Count, Microbial; D003470:Culture Media; D004271:DNA, Fungal / Q000032:analysis; D004064:Digestive System / Q000382:microbiology; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009503:Neutropenia / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D003470:Culture Media; D004271:DNA, Fungal