Molecular identification, antifungal resistance and virulence of Cryptococcus neoformans and Cryptococcus deneoformans isolated in Seville, Spain
Summary Cryptococcal meningitis is one of the leading causes of death in HIV/AIDS patients. Our aim was to in order to characterise the epidemiology, antifungal susceptibility pattern and virulence of 28 Cyptococcus sp. strains recovered from 12 AIDS patients during two years in a Spanish single ins...
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| Veröffentlicht in: | Mycoses 2017-01, Vol.60 (1), p.40-50 |
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| creator | Gago, Sara Serrano, Carmen Alastruey‐Izquierdo, Ana Cuesta, Isabel Martín‐Mazuelos, Estrella Aller, Ana Isabel Gómez‐López, Alicia Mellado, Emilia |
| description | Summary
Cryptococcal meningitis is one of the leading causes of death in HIV/AIDS patients. Our aim was to in order to characterise the epidemiology, antifungal susceptibility pattern and virulence of 28 Cyptococcus sp. strains recovered from 12 AIDS patients during two years in a Spanish single institution. Antifungal susceptibility testing was performed according to the CLSI protocols. Clinical strains were molecularly characterised by serotyping, mating type, PCR fingerprinting (M13 and GACA4 microsatellites) and analysis of two rDNA regions (IGS1 and ITS). Sequencing of the ERG11 gene was used to explore mechanisms of fluconazole resistance. Differences in virulence between species were studied in a Galleria mellonella infection model. Cryptococcus deneoformans and C. deneoformans x Cryptococcus neoformans hybrids were the most frequent variety (65%) followed by C. neoformans (35%). Strains were categorised according to 13 microsatellite genotypes and mixed infections could be detected in three patients. Twenty‐nine per cent of the strains were fluconazole resistant. In one of the patients, the fluconazole resistance phenotype was associated with a point mutation in the ERG11 gene responsible for the amino acid substitution G470R. C. neoformans strains were able to kill G. mellonella larvae more efficiently than C. deneoformans and hybrids between both species. Precisely molecular characterisation of C. neoformans species is important for an accurate patient's management. |
| doi_str_mv | 10.1111/myc.12543 |
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Cryptococcal meningitis is one of the leading causes of death in HIV/AIDS patients. Our aim was to in order to characterise the epidemiology, antifungal susceptibility pattern and virulence of 28 Cyptococcus sp. strains recovered from 12 AIDS patients during two years in a Spanish single institution. Antifungal susceptibility testing was performed according to the CLSI protocols. Clinical strains were molecularly characterised by serotyping, mating type, PCR fingerprinting (M13 and GACA4 microsatellites) and analysis of two rDNA regions (IGS1 and ITS). Sequencing of the ERG11 gene was used to explore mechanisms of fluconazole resistance. Differences in virulence between species were studied in a Galleria mellonella infection model. Cryptococcus deneoformans and C. deneoformans x Cryptococcus neoformans hybrids were the most frequent variety (65%) followed by C. neoformans (35%). Strains were categorised according to 13 microsatellite genotypes and mixed infections could be detected in three patients. Twenty‐nine per cent of the strains were fluconazole resistant. In one of the patients, the fluconazole resistance phenotype was associated with a point mutation in the ERG11 gene responsible for the amino acid substitution G470R. C. neoformans strains were able to kill G. mellonella larvae more efficiently than C. deneoformans and hybrids between both species. Precisely molecular characterisation of C. neoformans species is important for an accurate patient's management.</description><identifier>ISSN: 0933-7407</identifier><identifier>EISSN: 1439-0507</identifier><identifier>DOI: 10.1111/myc.12543</identifier><identifier>PMID: 27633849</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>Acquired immune deficiency syndrome ; Acquired Immunodeficiency Syndrome - complications ; Acquired Immunodeficiency Syndrome - microbiology ; AIDS ; Amino acid substitution ; Animals ; Antifungal Agents - pharmacology ; azole resistance evolution ; Cryptococcosis - drug therapy ; Cryptococcosis - epidemiology ; Cryptococcosis - microbiology ; Cryptococcus - drug effects ; Cryptococcus - genetics ; Cryptococcus - isolation & purification ; Cryptococcus - pathogenicity ; Cryptococcus neoformans ; Cryptococcus neoformans - drug effects ; Cryptococcus neoformans - genetics ; Cryptococcus neoformans - isolation & purification ; Cryptococcus neoformans - pathogenicity ; Cryptococcus species ; DNA Fingerprinting ; DNA, Fungal - chemistry ; DNA, Fungal - isolation & purification ; DNA, Intergenic - chemistry ; DNA, Intergenic - genetics ; Drug Resistance, Multiple, Fungal - genetics ; ERG11 gene ; Fingerprinting ; Fluconazole ; Fluconazole - pharmacology ; Galleria mellonella ; Genotypes ; Humans ; Hybrids ; Larva - microbiology ; Lentivirus ; Meningitis ; Meningitis, Cryptococcal - drug therapy ; Meningitis, Cryptococcal - epidemiology ; Meningitis, Cryptococcal - microbiology ; Microsatellites ; mixed infection ; Molecular Typing ; Moths - microbiology ; Mutation ; Point mutation ; Polymerase Chain Reaction ; Retroviridae ; Sequence Analysis, DNA ; Serotyping ; Spain - epidemiology ; Species ; Strains (organisms) ; Virulence</subject><ispartof>Mycoses, 2017-01, Vol.60 (1), p.40-50</ispartof><rights>2016 Blackwell Verlag GmbH</rights><rights>2016 Blackwell Verlag GmbH.</rights><rights>2017 Blackwell Verlag GmbH</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3863-8b96c9663194a22b03461460b0a2172c20e501a76176f6f150f63a328e292ddd3</citedby><cites>FETCH-LOGICAL-c3863-8b96c9663194a22b03461460b0a2172c20e501a76176f6f150f63a328e292ddd3</cites><orcidid>0000-0001-8651-4405</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fmyc.12543$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fmyc.12543$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27633849$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gago, Sara</creatorcontrib><creatorcontrib>Serrano, Carmen</creatorcontrib><creatorcontrib>Alastruey‐Izquierdo, Ana</creatorcontrib><creatorcontrib>Cuesta, Isabel</creatorcontrib><creatorcontrib>Martín‐Mazuelos, Estrella</creatorcontrib><creatorcontrib>Aller, Ana Isabel</creatorcontrib><creatorcontrib>Gómez‐López, Alicia</creatorcontrib><creatorcontrib>Mellado, Emilia</creatorcontrib><title>Molecular identification, antifungal resistance and virulence of Cryptococcus neoformans and Cryptococcus deneoformans isolated in Seville, Spain</title><title>Mycoses</title><addtitle>Mycoses</addtitle><description>Summary
Cryptococcal meningitis is one of the leading causes of death in HIV/AIDS patients. Our aim was to in order to characterise the epidemiology, antifungal susceptibility pattern and virulence of 28 Cyptococcus sp. strains recovered from 12 AIDS patients during two years in a Spanish single institution. Antifungal susceptibility testing was performed according to the CLSI protocols. Clinical strains were molecularly characterised by serotyping, mating type, PCR fingerprinting (M13 and GACA4 microsatellites) and analysis of two rDNA regions (IGS1 and ITS). Sequencing of the ERG11 gene was used to explore mechanisms of fluconazole resistance. Differences in virulence between species were studied in a Galleria mellonella infection model. Cryptococcus deneoformans and C. deneoformans x Cryptococcus neoformans hybrids were the most frequent variety (65%) followed by C. neoformans (35%). Strains were categorised according to 13 microsatellite genotypes and mixed infections could be detected in three patients. Twenty‐nine per cent of the strains were fluconazole resistant. In one of the patients, the fluconazole resistance phenotype was associated with a point mutation in the ERG11 gene responsible for the amino acid substitution G470R. C. neoformans strains were able to kill G. mellonella larvae more efficiently than C. deneoformans and hybrids between both species. Precisely molecular characterisation of C. neoformans species is important for an accurate patient's management.</description><subject>Acquired immune deficiency syndrome</subject><subject>Acquired Immunodeficiency Syndrome - complications</subject><subject>Acquired Immunodeficiency Syndrome - microbiology</subject><subject>AIDS</subject><subject>Amino acid substitution</subject><subject>Animals</subject><subject>Antifungal Agents - pharmacology</subject><subject>azole resistance evolution</subject><subject>Cryptococcosis - drug therapy</subject><subject>Cryptococcosis - epidemiology</subject><subject>Cryptococcosis - microbiology</subject><subject>Cryptococcus - drug effects</subject><subject>Cryptococcus - genetics</subject><subject>Cryptococcus - isolation & purification</subject><subject>Cryptococcus - pathogenicity</subject><subject>Cryptococcus neoformans</subject><subject>Cryptococcus neoformans - drug effects</subject><subject>Cryptococcus neoformans - genetics</subject><subject>Cryptococcus neoformans - isolation & purification</subject><subject>Cryptococcus neoformans - pathogenicity</subject><subject>Cryptococcus species</subject><subject>DNA Fingerprinting</subject><subject>DNA, Fungal - chemistry</subject><subject>DNA, Fungal - isolation & purification</subject><subject>DNA, Intergenic - chemistry</subject><subject>DNA, Intergenic - genetics</subject><subject>Drug Resistance, Multiple, Fungal - genetics</subject><subject>ERG11 gene</subject><subject>Fingerprinting</subject><subject>Fluconazole</subject><subject>Fluconazole - pharmacology</subject><subject>Galleria mellonella</subject><subject>Genotypes</subject><subject>Humans</subject><subject>Hybrids</subject><subject>Larva - microbiology</subject><subject>Lentivirus</subject><subject>Meningitis</subject><subject>Meningitis, Cryptococcal - drug therapy</subject><subject>Meningitis, Cryptococcal - epidemiology</subject><subject>Meningitis, Cryptococcal - microbiology</subject><subject>Microsatellites</subject><subject>mixed infection</subject><subject>Molecular Typing</subject><subject>Moths - microbiology</subject><subject>Mutation</subject><subject>Point mutation</subject><subject>Polymerase Chain Reaction</subject><subject>Retroviridae</subject><subject>Sequence Analysis, DNA</subject><subject>Serotyping</subject><subject>Spain - epidemiology</subject><subject>Species</subject><subject>Strains (organisms)</subject><subject>Virulence</subject><issn>0933-7407</issn><issn>1439-0507</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc-KFDEQh4Mo7jh68AUk4EVhezdJpdOdowz-g108rB48hUy6WrKkkzHpXpnH8I3N7KyKgmAuoVIfH1X5EfKUszNez_m0d2dctBLukRWXoBvWsu4-WTEN0HSSdSfkUSnXjPFOC_WQnIhOAfRSr8j3yxTQLcFm6geMsx-9s7NP8ZTaQ7XELzbQjMWX2UaH9XWgNz4vAQ9VGukm73dzcsm5pdCIaUx5srHcgn_0qv5315cU7IwD9ZFe4Y0PAU_p1c76-Jg8GG0o-OTuXpNPb15_3LxrLj68fb95ddE46BU0_VYrp5UCrqUVYstAKi4V2zIreCecYNgybjvFOzWqkbdsVGBB9Ci0GIYB1uTF0bvL6euCZTaTLw5DsHXMpRjet_UXoef8P1CpQEndQkWf_4VepyXHuojhWrAWelanX5OXR8rlVErG0eyyn2zeG87MIVJTIzW3kVb22Z1x2U44_CJ_ZliB8yPwzQfc_9tkLj9vjsofCP-rnw</recordid><startdate>201701</startdate><enddate>201701</enddate><creator>Gago, Sara</creator><creator>Serrano, Carmen</creator><creator>Alastruey‐Izquierdo, Ana</creator><creator>Cuesta, Isabel</creator><creator>Martín‐Mazuelos, Estrella</creator><creator>Aller, Ana Isabel</creator><creator>Gómez‐López, Alicia</creator><creator>Mellado, Emilia</creator><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>M7N</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0001-8651-4405</orcidid></search><sort><creationdate>201701</creationdate><title>Molecular identification, antifungal resistance and virulence of Cryptococcus neoformans and Cryptococcus deneoformans isolated in Seville, Spain</title><author>Gago, Sara ; Serrano, Carmen ; Alastruey‐Izquierdo, Ana ; Cuesta, Isabel ; Martín‐Mazuelos, Estrella ; Aller, Ana Isabel ; Gómez‐López, Alicia ; Mellado, Emilia</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3863-8b96c9663194a22b03461460b0a2172c20e501a76176f6f150f63a328e292ddd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Acquired immune deficiency syndrome</topic><topic>Acquired Immunodeficiency Syndrome - complications</topic><topic>Acquired Immunodeficiency Syndrome - microbiology</topic><topic>AIDS</topic><topic>Amino acid substitution</topic><topic>Animals</topic><topic>Antifungal Agents - pharmacology</topic><topic>azole resistance evolution</topic><topic>Cryptococcosis - drug therapy</topic><topic>Cryptococcosis - epidemiology</topic><topic>Cryptococcosis - microbiology</topic><topic>Cryptococcus - drug effects</topic><topic>Cryptococcus - genetics</topic><topic>Cryptococcus - isolation & purification</topic><topic>Cryptococcus - pathogenicity</topic><topic>Cryptococcus neoformans</topic><topic>Cryptococcus neoformans - drug effects</topic><topic>Cryptococcus neoformans - genetics</topic><topic>Cryptococcus neoformans - isolation & purification</topic><topic>Cryptococcus neoformans - pathogenicity</topic><topic>Cryptococcus species</topic><topic>DNA Fingerprinting</topic><topic>DNA, Fungal - chemistry</topic><topic>DNA, Fungal - isolation & purification</topic><topic>DNA, Intergenic - chemistry</topic><topic>DNA, Intergenic - genetics</topic><topic>Drug Resistance, Multiple, Fungal - genetics</topic><topic>ERG11 gene</topic><topic>Fingerprinting</topic><topic>Fluconazole</topic><topic>Fluconazole - pharmacology</topic><topic>Galleria mellonella</topic><topic>Genotypes</topic><topic>Humans</topic><topic>Hybrids</topic><topic>Larva - microbiology</topic><topic>Lentivirus</topic><topic>Meningitis</topic><topic>Meningitis, Cryptococcal - drug therapy</topic><topic>Meningitis, Cryptococcal - epidemiology</topic><topic>Meningitis, Cryptococcal - microbiology</topic><topic>Microsatellites</topic><topic>mixed infection</topic><topic>Molecular Typing</topic><topic>Moths - microbiology</topic><topic>Mutation</topic><topic>Point mutation</topic><topic>Polymerase Chain Reaction</topic><topic>Retroviridae</topic><topic>Sequence Analysis, DNA</topic><topic>Serotyping</topic><topic>Spain - epidemiology</topic><topic>Species</topic><topic>Strains (organisms)</topic><topic>Virulence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gago, Sara</creatorcontrib><creatorcontrib>Serrano, Carmen</creatorcontrib><creatorcontrib>Alastruey‐Izquierdo, Ana</creatorcontrib><creatorcontrib>Cuesta, Isabel</creatorcontrib><creatorcontrib>Martín‐Mazuelos, Estrella</creatorcontrib><creatorcontrib>Aller, Ana Isabel</creatorcontrib><creatorcontrib>Gómez‐López, Alicia</creatorcontrib><creatorcontrib>Mellado, Emilia</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>Mycoses</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gago, Sara</au><au>Serrano, Carmen</au><au>Alastruey‐Izquierdo, Ana</au><au>Cuesta, Isabel</au><au>Martín‐Mazuelos, Estrella</au><au>Aller, Ana Isabel</au><au>Gómez‐López, Alicia</au><au>Mellado, Emilia</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular identification, antifungal resistance and virulence of Cryptococcus neoformans and Cryptococcus deneoformans isolated in Seville, Spain</atitle><jtitle>Mycoses</jtitle><addtitle>Mycoses</addtitle><date>2017-01</date><risdate>2017</risdate><volume>60</volume><issue>1</issue><spage>40</spage><epage>50</epage><pages>40-50</pages><issn>0933-7407</issn><eissn>1439-0507</eissn><abstract>Summary
Cryptococcal meningitis is one of the leading causes of death in HIV/AIDS patients. Our aim was to in order to characterise the epidemiology, antifungal susceptibility pattern and virulence of 28 Cyptococcus sp. strains recovered from 12 AIDS patients during two years in a Spanish single institution. Antifungal susceptibility testing was performed according to the CLSI protocols. Clinical strains were molecularly characterised by serotyping, mating type, PCR fingerprinting (M13 and GACA4 microsatellites) and analysis of two rDNA regions (IGS1 and ITS). Sequencing of the ERG11 gene was used to explore mechanisms of fluconazole resistance. Differences in virulence between species were studied in a Galleria mellonella infection model. Cryptococcus deneoformans and C. deneoformans x Cryptococcus neoformans hybrids were the most frequent variety (65%) followed by C. neoformans (35%). Strains were categorised according to 13 microsatellite genotypes and mixed infections could be detected in three patients. Twenty‐nine per cent of the strains were fluconazole resistant. In one of the patients, the fluconazole resistance phenotype was associated with a point mutation in the ERG11 gene responsible for the amino acid substitution G470R. C. neoformans strains were able to kill G. mellonella larvae more efficiently than C. deneoformans and hybrids between both species. Precisely molecular characterisation of C. neoformans species is important for an accurate patient's management.</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>27633849</pmid><doi>10.1111/myc.12543</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-8651-4405</orcidid></addata></record> |
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| subjects | Acquired immune deficiency syndrome Acquired Immunodeficiency Syndrome - complications Acquired Immunodeficiency Syndrome - microbiology AIDS Amino acid substitution Animals Antifungal Agents - pharmacology azole resistance evolution Cryptococcosis - drug therapy Cryptococcosis - epidemiology Cryptococcosis - microbiology Cryptococcus - drug effects Cryptococcus - genetics Cryptococcus - isolation & purification Cryptococcus - pathogenicity Cryptococcus neoformans Cryptococcus neoformans - drug effects Cryptococcus neoformans - genetics Cryptococcus neoformans - isolation & purification Cryptococcus neoformans - pathogenicity Cryptococcus species DNA Fingerprinting DNA, Fungal - chemistry DNA, Fungal - isolation & purification DNA, Intergenic - chemistry DNA, Intergenic - genetics Drug Resistance, Multiple, Fungal - genetics ERG11 gene Fingerprinting Fluconazole Fluconazole - pharmacology Galleria mellonella Genotypes Humans Hybrids Larva - microbiology Lentivirus Meningitis Meningitis, Cryptococcal - drug therapy Meningitis, Cryptococcal - epidemiology Meningitis, Cryptococcal - microbiology Microsatellites mixed infection Molecular Typing Moths - microbiology Mutation Point mutation Polymerase Chain Reaction Retroviridae Sequence Analysis, DNA Serotyping Spain - epidemiology Species Strains (organisms) Virulence |
| title | Molecular identification, antifungal resistance and virulence of Cryptococcus neoformans and Cryptococcus deneoformans isolated in Seville, Spain |
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