Quantitation of pulmonary fungal burden in Paracoccidioides brasiliensis-infected mice by real-time PCR

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 deter...

Full description

Bibliographic Details
Published in:Revista do Instituto de Medicina Tropical de São Paulo
Main Authors: Marcelo Vieira Costa, Taise Natali Landgraf, Priscila C. Corrêa, Igor Emiliano Lemos Souza, Fabrício Freitas Fernandes, Ademilson Panunto-Castelo
Format: Article in Journal/Newspaper
Language:English
Published: Universidade de São Paulo (USP) 2018
Subjects:
Paracoccidioides brasiliensis
Paracoccidioidomycosis
Quantitative PCR
gp43 gene
Experimental infection
Real-Time PCR
Arctic medicine. Tropical medicine
RC955-962
Infectious and parasitic diseases
RC109-216
Arctic
Online Access:https://doi.org/10.1590/s1678-9946201961002
https://doaj.org/article/d4c03b1773c744108e8f2a814204cae4
Description
Summary: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.

Similar Items