2024-03-29T04:54:11Zhttp://repisalud.isciii.es/oai/requestoai:repisalud.isciii.es:20.500.12105/116852022-07-18T11:06:08Zcom_20.500.12105_2060com_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_2061
00925njm 22002777a 4500
dc
Carvalho-Pereira, Joana
author
Fernandes, Filipa
author
Araújo, Ricardo
author
Springer, Jan
author
Loeffler, Juergen
author
Buitrago, Maria Jose
author
Pais, Célia
author
Sampaio, Paula
author
2020-11-23
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.
J Fungi (Basel). 2020 Nov 23;6(4):308.
http://hdl.handle.net/20.500.12105/11685
33238439
10.3390/jof6040308
2309-608X
Journal of fungi (Basel, Switzerland)
Aspergillus sp.
Candida sp.
Rhizopus arrhizus
Fungal infections
Molecular diagnosis
Multiplex PCR
Multiplex PCR Based Strategy for Detection of Fungal Pathogen DNA in Patients with Suspected Invasive Fungal Infections