Quantitation of pulmonary fungal burden in Paracoccidioides brasiliensis-infected mice by real-time PCR
| Autor: | Taise Natali Landgraf, Igor Emiliano Lemos de Souza, Ademilson Panunto-Castelo, Priscila C. Corrêa, Marcelo Vieira Costa, Fabrício Freitas Fernandes |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 030106 microbiology Colony Count Microbial Real-Time Polymerase Chain Reaction Brief Communication Paracoccidioides Microbiology Mice 03 medical and health sciences chemistry.chemical_compound Quantitative PCR TaqMan medicine Animals gp43 gene Experimental infection Lung Gene Paracoccidioides brasiliensis chemistry.chemical_classification Mice Inbred BALB C Lung Diseases Fungal biology Paracoccidioidomycosis Stem Cells Reproducibility of Results biology.organism_classification medicine.disease Disease Models Animal 030104 developmental biology Real-time polymerase chain reaction chemistry Real-Time PCR Glycoprotein DNA |
| Zdroj: | Revista do Instituto de Medicina Tropical de São Paulo, Volume: 61, Article number: e2, Published: 20 DEC 2018 Revista do Instituto de Medicina Tropical de São Paulo, Vol 61, Iss 0 (2018) Revista do Instituto de Medicina Tropical de São Paulo |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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