Quantitation of pulmonary fungal burden in Paracoccidioides brasiliensis-infected mice by real-time PCR.
Marcelo Vieira CostaTaise Natali LandgrafPriscila C CorrêaIgor Emiliano Lemos SouzaFabrício Freitas FernandesAdemilson Panunto-CasteloPublished in: Revista do Instituto de Medicina Tropical de Sao Paulo (2018)
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.
Keyphrases
- real time pcr
- high throughput
- high fat diet induced
- copy number
- genome wide
- high resolution
- ms ms
- pulmonary hypertension
- mass spectrometry
- genome wide identification
- single molecule
- cell free
- insulin resistance
- gene expression
- liquid chromatography
- heat shock protein
- adipose tissue
- cell wall
- high performance liquid chromatography
- risk factors
- single cell
- tandem mass spectrometry
- skeletal muscle
- dna methylation