Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil

Patricia Fernanda Herkert; Jacques F Meis; Gabriel Lucca de Oliveira Salvador; Renata Rodrigues Gomes; Vania Aparecida Vicente; Marisol Dominguez Muro; Rosangela Lameira Pinheiro; Arnaldo Lopes Colombo; Alexandre Vargas Schwarzbold; Carla Sakuma de Oliveira; Marcelo Simão Ferreira; Flávio Queiroz-Telles; Ferry Hagen

doi:10.1099/jmm.0.000698

Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil

Patricia Fernanda Herkert, Jacques F Meis, Gabriel Lucca de Oliveira Salvador, Renata Rodrigues Gomes, Vania Aparecida Vicente, Marisol Dominguez Muro, Rosangela Lameira Pinheiro, Arnaldo Lopes Colombo, Alexandre Vargas Schwarzbold, Carla Sakuma de Oliveira, Marcelo Simão Ferreira, Flávio Queiroz-Telles, Ferry Hagen

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

18 Citations (Scopus)

Abstract

PURPOSE: Cryptococcosis is acquired from the environment by the inhalation of Cryptococcus cells and may establish from an asymptomatic latent infection into pneumonia or meningoencephalitis. The genetic diversity of a Cryptococcus neoformans species complex has been investigated by several molecular tools, such as multi-locus sequence typing, amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism and microsatellite analysis. This study aimed to investigate the genotype distributions and antifungal susceptibility profiles of C. neoformans sensu lato isolates from southern Brazil.

METHODOLOGY: We studied 219 C. neoformans sensu lato isolates with mating- and serotyping, AFLP fingerprinting, microsatellite typing and antifungal susceptibility testing.Results/Key findings. Among the isolates, 136 (69 %) were from HIV-positive patients. Only C. neoformans mating-type α and serotype A were observed. AFLP fingerprinting analysis divided the isolates into AFLP1/VNI (n=172; 78.5 %), AFLP1A/VNII (n=19; 8.7 %), AFLP1B/VNII (n=4; 1.8 %) and a new AFLP pattern AFLP1C (n=23; 10.5 %). All isolates were susceptible to tested antifungals and no correlation between antifungal susceptibility and genotypes was observed. Through microsatellite analysis, most isolates clustered in a major microsatellite complex and Simpson's diversity index of this population was D=0.9856.

CONCLUSION: The majority of C. neoformans sensu stricto infections occurred in HIV-positive patients. C. neoformans AFLP1/VNI was the most frequent genotype and all antifungal drugs had high in vitro activity against this species. Microsatellite analyses showed a high genetic diversity within the regional C. neoformans sensu stricto population, and correlation between environmental and clinical isolates, as well as a temporal and geographic relationship.

Original language	English
Pages (from-to)	560-569
Number of pages	10
Journal	Journal of Medical Microbiology
Volume	67
Issue number	4
DOIs	https://doi.org/10.1099/jmm.0.000698
Publication status	Published - Apr 2018

Keywords

Amplified Fragment Length Polymorphism Analysis
Antifungal Agents/pharmacology
Brazil
Cryptococcosis/microbiology
Cryptococcus neoformans/classification
Drug Resistance, Fungal
Genotype
Humans
Microbial Sensitivity Tests
Multilocus Sequence Typing
Phylogeny
Polymorphism, Restriction Fragment Length

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10.1099/jmm.0.000698

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Herkert, P. F., Meis, J. F., Lucca de Oliveira Salvador, G., Rodrigues Gomes, R., Aparecida Vicente, V., Dominguez Muro, M., Lameira Pinheiro, R., Lopes Colombo, A., Vargas Schwarzbold, A., Sakuma de Oliveira, C., Simão Ferreira, M., Queiroz-Telles, F., & Hagen, F. (2018). Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil. Journal of Medical Microbiology, 67(4), 560-569. https://doi.org/10.1099/jmm.0.000698

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title = "Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil",

abstract = "PURPOSE: Cryptococcosis is acquired from the environment by the inhalation of Cryptococcus cells and may establish from an asymptomatic latent infection into pneumonia or meningoencephalitis. The genetic diversity of a Cryptococcus neoformans species complex has been investigated by several molecular tools, such as multi-locus sequence typing, amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism and microsatellite analysis. This study aimed to investigate the genotype distributions and antifungal susceptibility profiles of C. neoformans sensu lato isolates from southern Brazil.METHODOLOGY: We studied 219 C. neoformans sensu lato isolates with mating- and serotyping, AFLP fingerprinting, microsatellite typing and antifungal susceptibility testing.Results/Key findings. Among the isolates, 136 (69 %) were from HIV-positive patients. Only C. neoformans mating-type α and serotype A were observed. AFLP fingerprinting analysis divided the isolates into AFLP1/VNI (n=172; 78.5 %), AFLP1A/VNII (n=19; 8.7 %), AFLP1B/VNII (n=4; 1.8 %) and a new AFLP pattern AFLP1C (n=23; 10.5 %). All isolates were susceptible to tested antifungals and no correlation between antifungal susceptibility and genotypes was observed. Through microsatellite analysis, most isolates clustered in a major microsatellite complex and Simpson's diversity index of this population was D=0.9856.CONCLUSION: The majority of C. neoformans sensu stricto infections occurred in HIV-positive patients. C. neoformans AFLP1/VNI was the most frequent genotype and all antifungal drugs had high in vitro activity against this species. Microsatellite analyses showed a high genetic diversity within the regional C. neoformans sensu stricto population, and correlation between environmental and clinical isolates, as well as a temporal and geographic relationship.",

keywords = "Amplified Fragment Length Polymorphism Analysis, Antifungal Agents/pharmacology, Brazil, Cryptococcosis/microbiology, Cryptococcus neoformans/classification, Drug Resistance, Fungal, Genotype, Humans, Microbial Sensitivity Tests, Multilocus Sequence Typing, Phylogeny, Polymorphism, Restriction Fragment Length",

author = "Herkert, {Patricia Fernanda} and Meis, {Jacques F} and {Lucca de Oliveira Salvador}, Gabriel and {Rodrigues Gomes}, Renata and {Aparecida Vicente}, Vania and {Dominguez Muro}, Marisol and {Lameira Pinheiro}, Rosangela and {Lopes Colombo}, Arnaldo and {Vargas Schwarzbold}, Alexandre and {Sakuma de Oliveira}, Carla and {Sim{\~a}o Ferreira}, Marcelo and Fl{\'a}vio Queiroz-Telles and Ferry Hagen",

year = "2018",

month = apr,

doi = "10.1099/jmm.0.000698",

language = "English",

volume = "67",

pages = "560--569",

journal = "Journal of Medical Microbiology",

issn = "0022-2615",

publisher = "Society for General Microbiology",

number = "4",

}

Herkert, PF, Meis, JF, Lucca de Oliveira Salvador, G, Rodrigues Gomes, R, Aparecida Vicente, V, Dominguez Muro, M, Lameira Pinheiro, R, Lopes Colombo, A, Vargas Schwarzbold, A, Sakuma de Oliveira, C, Simão Ferreira, M, Queiroz-Telles, F & Hagen, F 2018, 'Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil', Journal of Medical Microbiology, vol. 67, no. 4, pp. 560-569. https://doi.org/10.1099/jmm.0.000698

Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil. / Herkert, Patricia Fernanda; Meis, Jacques F; Lucca de Oliveira Salvador, Gabriel et al.
In: Journal of Medical Microbiology, Vol. 67, No. 4, 04.2018, p. 560-569.

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

TY - JOUR

T1 - Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil

AU - Herkert, Patricia Fernanda

AU - Meis, Jacques F

AU - Lucca de Oliveira Salvador, Gabriel

AU - Rodrigues Gomes, Renata

AU - Aparecida Vicente, Vania

AU - Dominguez Muro, Marisol

AU - Lameira Pinheiro, Rosangela

AU - Lopes Colombo, Arnaldo

AU - Vargas Schwarzbold, Alexandre

AU - Sakuma de Oliveira, Carla

AU - Simão Ferreira, Marcelo

AU - Queiroz-Telles, Flávio

AU - Hagen, Ferry

PY - 2018/4

Y1 - 2018/4

N2 - PURPOSE: Cryptococcosis is acquired from the environment by the inhalation of Cryptococcus cells and may establish from an asymptomatic latent infection into pneumonia or meningoencephalitis. The genetic diversity of a Cryptococcus neoformans species complex has been investigated by several molecular tools, such as multi-locus sequence typing, amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism and microsatellite analysis. This study aimed to investigate the genotype distributions and antifungal susceptibility profiles of C. neoformans sensu lato isolates from southern Brazil.METHODOLOGY: We studied 219 C. neoformans sensu lato isolates with mating- and serotyping, AFLP fingerprinting, microsatellite typing and antifungal susceptibility testing.Results/Key findings. Among the isolates, 136 (69 %) were from HIV-positive patients. Only C. neoformans mating-type α and serotype A were observed. AFLP fingerprinting analysis divided the isolates into AFLP1/VNI (n=172; 78.5 %), AFLP1A/VNII (n=19; 8.7 %), AFLP1B/VNII (n=4; 1.8 %) and a new AFLP pattern AFLP1C (n=23; 10.5 %). All isolates were susceptible to tested antifungals and no correlation between antifungal susceptibility and genotypes was observed. Through microsatellite analysis, most isolates clustered in a major microsatellite complex and Simpson's diversity index of this population was D=0.9856.CONCLUSION: The majority of C. neoformans sensu stricto infections occurred in HIV-positive patients. C. neoformans AFLP1/VNI was the most frequent genotype and all antifungal drugs had high in vitro activity against this species. Microsatellite analyses showed a high genetic diversity within the regional C. neoformans sensu stricto population, and correlation between environmental and clinical isolates, as well as a temporal and geographic relationship.

AB - PURPOSE: Cryptococcosis is acquired from the environment by the inhalation of Cryptococcus cells and may establish from an asymptomatic latent infection into pneumonia or meningoencephalitis. The genetic diversity of a Cryptococcus neoformans species complex has been investigated by several molecular tools, such as multi-locus sequence typing, amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism and microsatellite analysis. This study aimed to investigate the genotype distributions and antifungal susceptibility profiles of C. neoformans sensu lato isolates from southern Brazil.METHODOLOGY: We studied 219 C. neoformans sensu lato isolates with mating- and serotyping, AFLP fingerprinting, microsatellite typing and antifungal susceptibility testing.Results/Key findings. Among the isolates, 136 (69 %) were from HIV-positive patients. Only C. neoformans mating-type α and serotype A were observed. AFLP fingerprinting analysis divided the isolates into AFLP1/VNI (n=172; 78.5 %), AFLP1A/VNII (n=19; 8.7 %), AFLP1B/VNII (n=4; 1.8 %) and a new AFLP pattern AFLP1C (n=23; 10.5 %). All isolates were susceptible to tested antifungals and no correlation between antifungal susceptibility and genotypes was observed. Through microsatellite analysis, most isolates clustered in a major microsatellite complex and Simpson's diversity index of this population was D=0.9856.CONCLUSION: The majority of C. neoformans sensu stricto infections occurred in HIV-positive patients. C. neoformans AFLP1/VNI was the most frequent genotype and all antifungal drugs had high in vitro activity against this species. Microsatellite analyses showed a high genetic diversity within the regional C. neoformans sensu stricto population, and correlation between environmental and clinical isolates, as well as a temporal and geographic relationship.

KW - Amplified Fragment Length Polymorphism Analysis

KW - Antifungal Agents/pharmacology

KW - Brazil

KW - Cryptococcosis/microbiology

KW - Cryptococcus neoformans/classification

KW - Drug Resistance, Fungal

KW - Genotype

KW - Humans

KW - Microbial Sensitivity Tests

KW - Multilocus Sequence Typing

KW - Phylogeny

KW - Polymorphism, Restriction Fragment Length

U2 - 10.1099/jmm.0.000698

DO - 10.1099/jmm.0.000698

M3 - Article

C2 - 29461182

SN - 0022-2615

VL - 67

SP - 560

EP - 569

JO - Journal of Medical Microbiology

JF - Journal of Medical Microbiology

IS - 4

ER -

Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil

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Keywords

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