A flow cytometry method for testing the susceptibility of Cryptococcus spp. to amphotericin B

Benaducci, Tatiane; Matsumoto, Marcelo Teruyuki; Sardi, Janaína de Cássia Orlandi; Fusco‐Almeida, Ana Marisa; Mendes-Giannini, María José Soares

doi:10.1016/j.riam.2014.06.004

Cited by 11 publications

(9 citation statements)

References 29 publications

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“… Teodoro et al (2013) Susceptibility profile of clinical and environmental isolates of C. neoformans and C. gattii Andrade-Silva et al (2013) A flow cytometry method for testing the susceptibility of Cryptococcus spp. to AmB Benaducci et al (2015) In vitro susceptibility testing of AmB for C. neoformans variety grubii AFLP1/VNI and C. gattii AFLP6/VGII by CLSI and flow cytometry Morales et al (2015) Heteroresistance to ITZ and changes in the morphology and virulence of C. gattii Ferreira et al (2015) Antifungal susceptibility of clinical C. deuterogattii (AFLP6/VGII) isolates from Southern Brazil Herkert et al (2016) Resistance to FCZ and changes on virulence and morphological aspects of C. neoformans and C. gattii Rossi et al (2016) Antifungal susceptibility testing and genotyping characterization of C. neoformans and C. gattii isolates from HIV-infected patients Figueiredo et al (2016) Evaluation of antifungal combination against Cryptococcus spp. Reichert-Lima et al (2016) C. neoformans and C. gattii isolates from both HIV-infected and uninfected patients: antifungal susceptibility and outcome of cryptococcal disease Nascimento et al (2017) FCZ non-susceptible C. neoformans .…”

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“…Several additional techniques have been developed to simplify the standard ones, such as, time kill curves (TKC), E-Test strips, and a disk diffusion method ( Ochiuzzi et al 2010 , Córdoba et al 2015 , de Oliveira et al 2017 ). Recently, the use of flow cytometry (FC) was evaluated in two publications, which both showed a positive significant correlation ( Benaducci et al 2015 , Morales et al 2015 ). In one of them, the conclusion was that FC has an excellent correlation with the standard methods, and it is a reliable, fast, and safe method to test the susceptibility to AmB ( Benaducci et al 2015 ).…”

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“…Recently, the use of flow cytometry (FC) was evaluated in two publications, which both showed a positive significant correlation ( Benaducci et al 2015 , Morales et al 2015 ). In one of them, the conclusion was that FC has an excellent correlation with the standard methods, and it is a reliable, fast, and safe method to test the susceptibility to AmB ( Benaducci et al 2015 ). In the other study, the antifungal susceptibility of 17 clinical isolates of C. neoformans and 18 C. gattii VGII, was analysed by both the microdilution method (CLSI M27-A3) and FC, showing that FC allowed to determine MIC for AmB in a 2 h incubation time, with MICs data obtained on the same day ( Morales et al 2015 ).…”

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The status of cryptococcosis in Latin America

Firacative

Lizarazo

Illnait-Zaragozí

et al. 2018

Mem. Inst. Oswaldo Cruz

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Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America.

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The status of cryptococcosis in Latin America

Firacative

Lizarazo

Illnait-Zaragozí

et al. 2018

Mem. Inst. Oswaldo Cruz

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“…Amplification of DNA-extracted blood samples is hampered by the presence of hemoglobin and anticoagulants [35,36]. Some DNA extraction businesses address this issue by incorporating treatment stages to eliminate potential inhibitors, which can be a problem with other methods.…”

Section: End-point Pcr-based Amplificationmentioning

confidence: 99%

Molecular Detection and Identification of Candida

Al-Taee¹

2023

Candida and Candidiasis

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Human opportunistic yeast infections have become more common in recent years. Many infections are difficult to treat and diagnose due to the large number and diversity of organisms that can cause sickness. In addition, infectious strains eventually develop resistance to one or more antifungal medicines, severely limiting treatment choices and emphasizing the need of early detection of the infective agent and its drug sensitivity profile. Current techniques for detecting species and resistances are insensitive and specific, and they frequently need pre-cultivation of the causal agent, which delays diagnosis. New high-throughput technologies, such as next-generation sequencing or proteomics, make it possible to identify yeast infections more sensitively, accurately, and quickly. Opportunistic yeast pathogens, cause a wide spectrum of superficial and systemic infections, many of which are lethal. In this work, we give an overview of current and newly created approaches. It may be used to determine the presence of yeast infections as well as their medication resistance. Throughout the book, we highlight the following points: Explaining the benefits and drawbacks of each strategy, as well as the most promising advancements on their route to success.

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“…Pore et al first proposed PI and rose Bengal to analyze cell viability in the context of AFST (Pore, 1990). Since these promising results, many studies have used PI but further results were less conclusive depending on the drug and the experimental protocols (Pore, 1991;Green et al, 1994;Ramani and Chaturvedi, 2000;Chaturvedi et al, 2004;Benaducci et al, 2015). Besides PI, the two most promising fluorescent probes in AFST are AO and FUN-1.…”

Section: Principle and Dyesmentioning

confidence: 99%

Can We Improve Antifungal Susceptibility Testing?

Durand

Maubon

Cornet

et al. 2021

Front. Cell. Infect. Microbiol.

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Systemic antifungal agents are increasingly used for prevention or treatment of invasive fungal infections, whose prognosis remains poor. At the same time, emergence of resistant or even multi-resistant strains is of concern as the antifungal arsenal is limited. Antifungal susceptibility testing (AFST) is therefore of key importance for patient management and antifungal stewardship. Current AFST methods, including reference and commercial types, are based on growth inhibition in the presence of an antifungal, in liquid or solid media. They usually enable Minimal Inhibitory Concentrations (MIC) to be determined with direct clinical application. However, they are limited by a high turnaround time (TAT). Several innovative methods are currently under development to improve AFST. Techniques based on MALDI-TOF are promising with short TAT, but still need extensive clinical validation. Flow cytometry and computed imaging techniques detecting cellular responses to antifungal stress other than growth inhibition are also of interest. Finally, molecular detection of mutations associated with antifungal resistance is an intriguing alternative to standard AFST, already used in routine microbiology labs for detection of azole resistance in Aspergillus and even directly from samples. It is still restricted to known mutations. The development of Next Generation Sequencing (NGS) and whole-genome approaches may overcome this limitation in the near future. While promising approaches are under development, they are not perfect and the ideal AFST technique (user-friendly, reproducible, low-cost, fast and accurate) still needs to be set up routinely in clinical laboratories.

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A flow cytometry method for testing the susceptibility of Cryptococcus spp. to amphotericin B

Cited by 11 publications

References 29 publications

The status of cryptococcosis in Latin America

The status of cryptococcosis in Latin America

Molecular Detection and Identification of Candida

Can We Improve Antifungal Susceptibility Testing?

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