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

Costa, Marcelo Vieira; Landgraf, Taise Natali; Corrêa, Priscila Cristina; Souza, Igor Emiliano Lemos de; Fernandes, Fabrício Freitas; Panunto-Castelo, Ademílson

doi:10.1590/s1678-9946201961002

Cited by 3 publications

(4 citation statements)

References 16 publications

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“…The assay was later used to detect Paracoccidioides DNA in tissue and peripheral blood of a patient that presented positive serology but negative culture [ 124 ]. Another interesting application of this technique was quantifying fungal load in experimentally infected mice [ 125 ]. Though promising, the method cannot differentiate between living and dead cells, which can be a problem [ 125 ].…”

Section: Molecular Diagnosis: Where Do We Stand?mentioning

confidence: 99%

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Molecular Tools for Detection and Identification of Paracoccidioides Species: Current Status and Future Perspectives

Pinheiro

Hahn

Camargo

et al. 2020

JoF

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Paracoccidioidomycosis (PCM) is a mycotic disease caused by the Paracoccidioides species, a group of thermally dimorphic fungi that grow in mycelial form at 25 °C and as budding yeasts when cultured at 37 °C or when parasitizing the host tissues. PCM occurs in a large area of Latin America, and the most critical regions of endemicity are in Brazil, Colombia, and Venezuela. The clinical diagnosis of PCM needs to be confirmed through laboratory tests. Although classical laboratory techniques provide valuable information due to the presence of pathognomonic forms of Paracoccidioides spp., nucleic acid-based diagnostics gradually are replacing or complementing culture-based, biochemical, and immunological assays in routine microbiology laboratory practice. Recently, taxonomic changes driven by whole-genomic sequencing of Paracoccidioides have highlighted the need to recognize species boundaries, which could better ascertain Paracoccidioides taxonomy. In this scenario, classical laboratory techniques do not have significant discriminatory power over cryptic agents. On the other hand, several PCR-based methods can detect polymorphisms in Paracoccidioides DNA and thus support species identification. This review is focused on the recent achievements in molecular diagnostics of paracoccidioidomycosis, including the main advantages and pitfalls related to each technique. We discuss these breakthroughs in light of taxonomic changes in the Paracoccidioides genus.

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Section: Molecular Diagnosis: Where Do We Stand?mentioning

confidence: 99%

“…Another interesting application of this technique was quantifying fungal load in experimentally infected mice [ 125 ]. Though promising, the method cannot differentiate between living and dead cells, which can be a problem [ 125 ].…”

Section: Molecular Diagnosis: Where Do We Stand?mentioning

confidence: 99%

Molecular Tools for Detection and Identification of Paracoccidioides Species: Current Status and Future Perspectives

Pinheiro

Hahn

Camargo

et al. 2020

JoF

View full text Add to dashboard Cite

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“…For molecular detection of fungal pathogens, PCR is the most preferred method [215,216] and is regarded as a standard platform in many clinical laboratories, even in developing countries, due to its affordability and reproducibility [217,218]. RT-PCR assays with a short turnaround time can provide desirable alternatives for the rapid detection of microbes [210,219], and they are able to quantify the amount of amplified DNA in real time. Hence, conventional PCR methods have been replaced by RT-PCR techniques in medical laboratories [216].…”

Section: Molecular Methods Used To Detect Yeastsmentioning

confidence: 99%

“…SYBR ® Green- and TaqMan-based assays using primers and probe for the 43-kDa glycoprotein (gp43) gene were able to detect as little as 270 gene copies (about 2 fg of DNA) per reaction. Although qPCR assays cannot distinguish between living and dead yeasts, it found a highly positive linear correlation between CFU and qPCR [219]. PCR and RT-PCR assays were also used to 100% identify 44 C. auris and related species, such as Candida duobushaemulonii , C. haemulonii , and C. lusitaniae with strong results [232].…”

Section: Molecular Methods Used To Detect Yeastsmentioning

confidence: 99%

Advances in Chemical and Biological Methods to Identify Microorganisms—From Past to Present

et al. 2019

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Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal isolates and current analysis call for broad reports of microorganisms, involving the application of molecular techniques (e.g., 16S ribosomal RNA gene sequencing based on polymerase chain reaction). The goal of this review is to present the past and the present methods of detection and identification of microorganisms, and to discuss their advantages and their limitations.

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Paracoccidioidomycosis: Current Status and Future Trends

et al. 2022

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Paracoccidioidomycosis (PCM), initially reported in 1908 in the city of São Paulo, Brazil, by Adolpho Lutz, is primarily a systemic and neglected tropical mycosis that may affect individuals with certain risk factors around Latin America, especially Brazil. Paracoccidioides brasiliensis sensu stricto , a classical thermodimorphic fungus associated with PCM, was long considered to represent a monotypic taxon. However, advances in molecular taxonomy revealed several cryptic species, including Paracoccidioides americana , P. restrepiensis , P. venezuelensis , and P. lutzii , that show a preference for skin and mucous membranes, lymph nodes, and respiratory organs but can also affect many other organs.

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

Cited by 3 publications

References 16 publications

Molecular Tools for Detection and Identification of Paracoccidioides Species: Current Status and Future Perspectives

Molecular Tools for Detection and Identification of Paracoccidioides Species: Current Status and Future Perspectives

Advances in Chemical and Biological Methods to Identify Microorganisms—From Past to Present

Paracoccidioidomycosis: Current Status and Future Trends

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