title: MLST-Based Population Genetic Analysis in a Global Context Reveals Clonality amongst Cryptococcus neoformans var. grubii VNI Isolates from HIV Patients in Southeastern Brazil.
abstract: Cryptococcosis is an important fungal infection in immunocompromised individuals, especially those infected with HIV. In Brazil, despite the free availability of antiretroviral therapy (ART) in the public health system, the mortality rate due to Cryptococcus neoformans meningitis is still high. To obtain a more detailed picture of the population genetic structure of this species in southeast Brazil, we studied 108 clinical isolates from 101 patients and 35 environmental isolates. Among the patients, 59% had a fatal outcome mainly in HIV-positive male patients. All the isolates were found to be C. neoformans var. grubii major molecular type VNI and mating type locus alpha. Twelve were identified as diploid by flow cytometry, being homozygous (AαAα) for the mating type and by PCR screening of the STE20, GPA1, and PAK1 genes. Using the ISHAM consensus multilocus sequence typing (MLST) scheme, 13 sequence types (ST) were identified, with one being newly described. ST93 was identified from 81 (75%) of the clinical isolates, while ST77 and ST93 were identified from 19 (54%) and 10 (29%) environmental isolates, respectively. The southeastern Brazilian isolates had an overwhelming clonal population structure. When compared with populations from different continents based on data extracted from the ISHAM-MLST database (mlst.mycologylab.org) they showed less genetic variability. Two main clusters within C. neoformans var. grubii VNI were identified that diverged from VNB around 0.58 to 4.8 million years ago.
authors.affiliation: Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Marie Bashir Institute for Emerging Infectious Diseases and Biosecurity, Sydney Medical School-Westmead Hospital, The Westmead Institute for Medical Research, The University of Sydney, Sydney, Australia.|Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Kennio
authors.initials: K
authors.lastname: Ferreira-Paim
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Andrade-Silva
authors.affiliation: Biomedicine Department, Federal University of Piauí, Parnaíba, Brazil.
authors.forename: Fernanda M
authors.initials: FM
authors.lastname: Fonseca
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Thatiana B
authors.initials: TB
authors.lastname: Ferreira
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Delio J
authors.initials: DJ
authors.lastname: Mora
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: Andrade-Silva
authors.affiliation: Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Marie Bashir Institute for Emerging Infectious Diseases and Biosecurity, Sydney Medical School-Westmead Hospital, The Westmead Institute for Medical Research, The University of Sydney, Sydney, Australia.
authors.forename: Aziza
authors.initials: A
authors.lastname: Khan
authors.affiliation: Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Marie Bashir Institute for Emerging Infectious Diseases and Biosecurity, Sydney Medical School-Westmead Hospital, The Westmead Institute for Medical Research, The University of Sydney, Sydney, Australia.
authors.forename: Aiken
authors.initials: A
authors.lastname: Dao
authors.affiliation: Infectious Disease Department, Faculty of Medicine of São José do Rio Preto, São José do Rio Preto, Brazil.
authors.forename: Eduardo C
authors.initials: EC
authors.lastname: Reis
authors.affiliation: Infectious Disease Department, Faculty of Medicine of São José do Rio Preto, São José do Rio Preto, Brazil.
authors.forename: Margarete T G
authors.initials: MT
authors.lastname: Almeida
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Andre
authors.initials: A
authors.lastname: Maltos
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Virmondes R
authors.initials: VR
authors.lastname: Junior
authors.affiliation: Evandro Chagas National Institute of Infectious Diseases, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
authors.forename: Luciana
authors.initials: L
authors.lastname: Trilles
authors.affiliation: Mycology Section, Robert Koch Institute, Berlin, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Rickerts
authors.affiliation: Mycology Unit, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Ariya
authors.initials: A
authors.lastname: Chindamporn
authors.affiliation: Department of Medicine and Epidemiology, University of California, Davis, United States of America.
authors.forename: Jane E
authors.initials: JE
authors.lastname: Sykes
authors.affiliation: Laboratorio Micologia Medica, Dip. Scienze Biomediche per la Salute, Università degli Studi di Milano, Milano, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Cogliati
authors.affiliation: Department of Microbiology and Immunology, Medical School, University of Minnesota, Minneapolis, Mississippi, United States of America.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Nielsen
authors.affiliation: Department of Yeast and Basidiomycete Research, CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands.
authors.forename: Teun
authors.initials: T
authors.lastname: Boekhout
authors.affiliation: Department of Infectious Disease Epidemiology, Imperial College London, Norfolk Place, London, United Kingdom.
authors.forename: Matthew
authors.initials: M
authors.lastname: Fisher
authors.affiliation: Molecular Microbiology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institute of Health, Bethesda, Maryland, United States of America.
authors.forename: June
authors.initials: J
authors.lastname: Kwon-Chung
authors.affiliation: Translational Genomics Research Institute, Flagstaff, Arizona, United States of America.
authors.forename: David M
authors.initials: DM
authors.lastname: Engelthaler
authors.affiliation: Evandro Chagas National Institute of Infectious Diseases, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
authors.forename: Marcia
authors.initials: M
authors.lastname: Lazéra
authors.affiliation: Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Marie Bashir Institute for Emerging Infectious Diseases and Biosecurity, Sydney Medical School-Westmead Hospital, The Westmead Institute for Medical Research, The University of Sydney, Sydney, Australia.
authors.forename: Wieland
authors.initials: W
authors.lastname: Meyer
authors.affiliation: Infectious Disease Department, Triangulo Mineiro Federal University, Uberaba, Brazil.
authors.forename: Mario L
authors.initials: ML
authors.lastname: Silva-Vergara
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000382:microbiology*; D000328:Adult; D001938:Brazil; D003453:Cryptococcosis / Q000209:etiology / Q000382:microbiology*; D003455:Cryptococcus neoformans / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D005260:Female; D005838:Genotype; D015658:HIV Infections / Q000150:complications* / Q000821:virology; D006801:Humans; D008297:Male; D008875:Middle Aged; D058885:Multilocus Sequence Typing; D016533:Mycological Typing Techniques; D010802:Phylogeny; D055815:Young Adult