Quantitation of pulmonary fungal burden in Paracoccidioides brasiliensis-infected mice by real-time PCR
AUTOR(ES)
Costa, Marcelo Vieira, Landgraf, Taise Natali, Corrêa, Priscila C., Souza, Igor Emiliano Lemos, Fernandes, Fabrício Freitas, Panunto-Castelo, Ademilson
FONTE
Rev. Inst. Med. trop. S. Paulo
DATA DE PUBLICAÇÃO
20/12/2018
RESUMO
ABSTRACT Although colony-forming unit (CFU) counting is widely used to quantify fungal load in tissue from animal experimentally infected with Paracoccidioides brasiliensis, several technical disadvantages have been described. Here we developed highly accurate quantitative PCR (qPCR) assays to determine the relative P brasiliensis load in lungs from infected mice. SYBR Green- and TaqMan-based assays using primers and probe for the 43-kDa glycoprotein (gp43) gene detected as little as 270 gene copies (about 2 fg of DNA) per reaction. Although qPCR assays cannot distinguish between living and dead yeasts, we found a highly positive linear correlation between CFU and qPCR.
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