TY  - JOUR
A1  - Carvalho-Pereira, Joana
A1  - Fernandes, Filipa
A1  - Araújo, Ricardo
A1  - Springer, Jan
A1  - Loeffler, Juergen
A1  - Buitrago, María José
A1  - Pais, Célia
A1  - Sampaio, Paula
T1  - Multiplex PCR based strategy for detection of fungal pathogen DNA in patients with suspected invasive fungal infections
JF  - Journal of Fungi
N2  - A new and easy polymerase chain reaction (PCR) multiplex strategy, for the identification of the most common fungal species involved in invasive fungal infections (IFI) was developed in this work. Two panels with species-specific markers were designed, the Candida Panel for the identification of Candida species, and the Filamentous Fungi Panel for the identification of Aspergillus species and Rhizopusarrhizus. The method allowed the correct identification of all targeted pathogens using extracted DNA or by colony PCR, showed no cross-reactivity with nontargeted species and allowed identification of different species in mixed infections. Sensitivity reached 10 to 1 pg of DNA and was suitable for clinical samples from sterile sites, with a sensitivity of 89% and specificity of 100%. Overall, the study showed that the new method is suitable for the identification of the ten most important fungal species involved in IFI, not only from positive blood cultures but also from clinical samples from sterile sites. The method provides a unique characteristic, of seeing the peak in the specific region of the panel with the correct fluorescence dye, that aids the ruling out of unspecific amplifications. Furthermore, the panels can be further customized, selecting markers for different species and/or resistance genes.
KW  - molecular diagnosis
KW  - fungal infections
KW  - multiplex PCR
KW  - Candida sp.
KW  - Aspergillus sp.
KW  - Rhizopus arrhizus
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219392
SN  - 2309-608X
VL  - 6
IS  - 4
ER  -