Simple, Low-Cost Detection of Candida parapsilosis Complex Isolates and Molecular Fingerprinting of Candida orthopsilosis Strains in Kuwait by ITS Region Sequencing and Amplified Fragment Length Polymorphism Analysis

Candida parapsilosis has now emerged as the second or third most important cause of healthcare-associated Candida infections. Molecular studies have shown that phenotypically identified C. parapsilosis isolates represent a complex of three species, namely, C. parapsilosis, C. orthopsilosis and C. me...

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Veröffentlicht in:	PloS one Jg. 10; H. 11; S. e0142880
Hauptverfasser:	Asadzadeh, Mohammad, Ahmad, Suhail, Hagen, Ferry, Meis, Jacques F., Al-Sweih, Noura, Khan, Ziauddin
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	United States Public Library of Science 18.11.2015 Public Library of Science (PLoS)
Schlagworte:	Amplified fragment length polymorphism Analysis Antifungal Agents - pharmacology Candida Candida - classification Candida - drug effects Candida - genetics Candida - isolation & purification Candida albicans Candida parapsilosis Candidiasis Candidiasis - diagnosis Candidiasis - microbiology Care and treatment Chemical fingerprinting Clinical isolates Deoxyribonucleic acid Desorption Development and progression DNA DNA Fingerprinting - methods DNA sequencing DNA, Fungal - genetics DNA, Intergenic - genetics DNA, Ribosomal - genetics Economic aspects Epidemiology Fingerprinting Gene polymorphism Gene sequencing Genetic aspects Genomes Genotype Genotypes Haplotypes Health aspects Health care Hospitals Humans Identification Infections Infectious diseases Kuwait Lasers Low cost Mass spectrometry Medicine Microbial Sensitivity Tests Multiplex Polymerase Chain Reaction - economics Multiplex Polymerase Chain Reaction - methods Multiplexing Mycological Typing Techniques Pathogens Patient outcomes Patients Phylogeny Polymerase chain reaction Polymorphism Polymorphism, Restriction Fragment Length Scientific imaging Sequence Analysis, DNA Spacer Species Studies Yeast Kuwait Netherlands Germany
ISSN:	1932-6203, 1932-6203
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Abstract	Candida parapsilosis has now emerged as the second or third most important cause of healthcare-associated Candida infections. Molecular studies have shown that phenotypically identified C. parapsilosis isolates represent a complex of three species, namely, C. parapsilosis, C. orthopsilosis and C. metapsilosis. Lodderomyces elongisporus is another species phenotypically closely related to the C. parapsilosis-complex. The aim of this study was to develop a simple, low cost multiplex (m) PCR assay for species-specific identification of C. parapsilosis complex isolates and to study genetic relatedness of C. orthopsilosis isolates in Kuwait. Species-specific amplicons from C. parapsilosis (171 bp), C. orthopsilosis (109 bp), C. metapsilosis (217 bp) and L. elongisporus (258 bp) were obtained in mPCR. Clinical isolates identified as C. parapsilosis (n = 380) by Vitek2 in Kuwait and an international collection of 27 C. parapsilosis complex and L. elongisporus isolates previously characterized by rDNA sequencing were analyzed to evaluate mPCR. Species-specific PCR and DNA sequencing of internal transcribed spacer (ITS) region of rDNA were performed to validate the results of mPCR. Fingerprinting of 19 clinical C. orthopsilosis isolates (including 4 isolates from a previous study) was performed by amplified fragment length polymorphism (AFLP) analysis. Phenotypically identified C. parapsilosis isolates (n = 380) were identified as C. parapsilosis sensu stricto (n = 361), C. orthopsilosis (n = 15), C. metapsilosis (n = 1) and L. elongisporus (n = 3) by mPCR. The mPCR also accurately detected all epidemiologically unrelated C. parapsilosis complex and L. elongisporus isolates. The 19 C. orthopsilosis isolates obtained from 16 patients were divided into 3 haplotypes based on ITS region sequence data. Seven distinct genotypes were identified among the 19 C. orthopsilosis isolates by AFLP including a dominant genotype (AFLP1) comprising 11 isolates recovered from 10 patients. A rapid, low-cost mPCR assay for detection and differentiation of C. parapsilosis, C. orthopsilosis, C. metapsilosis and L. elongisporus has been developed.
AbstractList	Candida parapsilosis has now emerged as the second or third most important cause of healthcare-associated Candida infections. Molecular studies have shown that phenotypically identified C. parapsilosis isolates represent a complex of three species, namely, C. parapsilosis, C. orthopsilosis and C. metapsilosis. Lodderomyces elongisporus is another species phenotypically closely related to the C. parapsilosis-complex. The aim of this study was to develop a simple, low cost multiplex (m) PCR assay for species-specific identification of C. parapsilosis complex isolates and to study genetic relatedness of C. orthopsilosis isolates in Kuwait. Species-specific amplicons from C. parapsilosis (171 bp), C. orthopsilosis (109 bp), C. metapsilosis (217 bp) and L. elongisporus (258 bp) were obtained in mPCR. Clinical isolates identified as C. parapsilosis (n = 380) by Vitek2 in Kuwait and an international collection of 27 C. parapsilosis complex and L. elongisporus isolates previously characterized by rDNA sequencing were analyzed to evaluate mPCR. Species-specific PCR and DNA sequencing of internal transcribed spacer (ITS) region of rDNA were performed to validate the results of mPCR. Fingerprinting of 19 clinical C. orthopsilosis isolates (including 4 isolates from a previous study) was performed by amplified fragment length polymorphism (AFLP) analysis. Phenotypically identified C. parapsilosis isolates (n = 380) were identified as C. parapsilosis sensu stricto (n = 361), C. orthopsilosis (n = 15), C. metapsilosis (n = 1) and L. elongisporus (n = 3) by mPCR. The mPCR also accurately detected all epidemiologically unrelated C. parapsilosis complex and L. elongisporus isolates. The 19 C. orthopsilosis isolates obtained from 16 patients were divided into 3 haplotypes based on ITS region sequence data. Seven distinct genotypes were identified among the 19 C. orthopsilosis isolates by AFLP including a dominant genotype (AFLP1) comprising 11 isolates recovered from 10 patients. A rapid, low-cost mPCR assay for detection and differentiation of C. parapsilosis, C. orthopsilosis, C. metapsilosis and L. elongisporus has been developed. Candida parapsilosis has now emerged as the second or third most important cause of healthcare-associated Candida infections. Molecular studies have shown that phenotypically identified C. parapsilosis isolates represent a complex of three species, namely, C. parapsilosis, C. orthopsilosis and C. metapsilosis. Lodderomyces elongisporus is another species phenotypically closely related to the C. parapsilosis-complex. The aim of this study was to develop a simple, low cost multiplex (m) PCR assay for species-specific identification of C. parapsilosis complex isolates and to study genetic relatedness of C. orthopsilosis isolates in Kuwait. Species-specific amplicons from C. parapsilosis (171 bp), C. orthopsilosis (109 bp), C. metapsilosis (217 bp) and L. elongisporus (258 bp) were obtained in mPCR. Clinical isolates identified as C. parapsilosis (n = 380) by Vitek2 in Kuwait and an international collection of 27 C. parapsilosis complex and L. elongisporus isolates previously characterized by rDNA sequencing were analyzed to evaluate mPCR. Species-specific PCR and DNA sequencing of internal transcribed spacer (ITS) region of rDNA were performed to validate the results of mPCR. Fingerprinting of 19 clinical C. orthopsilosis isolates (including 4 isolates from a previous study) was performed by amplified fragment length polymorphism (AFLP) analysis. Phenotypically identified C. parapsilosis isolates (n = 380) were identified as C. parapsilosis sensu stricto (n = 361), C. orthopsilosis (n = 15), C. metapsilosis (n = 1) and L. elongisporus (n = 3) by mPCR. The mPCR also accurately detected all epidemiologically unrelated C. parapsilosis complex and L. elongisporus isolates. The 19 C. orthopsilosis isolates obtained from 16 patients were divided into 3 haplotypes based on ITS region sequence data. Seven distinct genotypes were identified among the 19 C. orthopsilosis isolates by AFLP including a dominant genotype (AFLP1) comprising 11 isolates recovered from 10 patients. A rapid, low-cost mPCR assay for detection and differentiation of C. parapsilosis, C. orthopsilosis, C. metapsilosis and L. elongisporus has been developed.Candida parapsilosis has now emerged as the second or third most important cause of healthcare-associated Candida infections. Molecular studies have shown that phenotypically identified C. parapsilosis isolates represent a complex of three species, namely, C. parapsilosis, C. orthopsilosis and C. metapsilosis. Lodderomyces elongisporus is another species phenotypically closely related to the C. parapsilosis-complex. The aim of this study was to develop a simple, low cost multiplex (m) PCR assay for species-specific identification of C. parapsilosis complex isolates and to study genetic relatedness of C. orthopsilosis isolates in Kuwait. Species-specific amplicons from C. parapsilosis (171 bp), C. orthopsilosis (109 bp), C. metapsilosis (217 bp) and L. elongisporus (258 bp) were obtained in mPCR. Clinical isolates identified as C. parapsilosis (n = 380) by Vitek2 in Kuwait and an international collection of 27 C. parapsilosis complex and L. elongisporus isolates previously characterized by rDNA sequencing were analyzed to evaluate mPCR. Species-specific PCR and DNA sequencing of internal transcribed spacer (ITS) region of rDNA were performed to validate the results of mPCR. Fingerprinting of 19 clinical C. orthopsilosis isolates (including 4 isolates from a previous study) was performed by amplified fragment length polymorphism (AFLP) analysis. Phenotypically identified C. parapsilosis isolates (n = 380) were identified as C. parapsilosis sensu stricto (n = 361), C. orthopsilosis (n = 15), C. metapsilosis (n = 1) and L. elongisporus (n = 3) by mPCR. The mPCR also accurately detected all epidemiologically unrelated C. parapsilosis complex and L. elongisporus isolates. The 19 C. orthopsilosis isolates obtained from 16 patients were divided into 3 haplotypes based on ITS region sequence data. Seven distinct genotypes were identified among the 19 C. orthopsilosis isolates by AFLP including a dominant genotype (AFLP1) comprising 11 isolates recovered from 10 patients. A rapid, low-cost mPCR assay for detection and differentiation of C. parapsilosis, C. orthopsilosis, C. metapsilosis and L. elongisporus has been developed.
Audience	Academic
Author	Al-Sweih, Noura Khan, Ziauddin Meis, Jacques F. Hagen, Ferry Asadzadeh, Mohammad Ahmad, Suhail
AuthorAffiliation	3 Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands California Department of Public Health, UNITED STATES 1 Department of Microbiology, Faculty of Medicine, Kuwait University, Safat, Kuwait 2 Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands
AuthorAffiliation_xml	– name: 1 Department of Microbiology, Faculty of Medicine, Kuwait University, Safat, Kuwait – name: 3 Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands – name: California Department of Public Health, UNITED STATES – name: 2 Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands
Author_xml	– sequence: 1 givenname: Mohammad surname: Asadzadeh fullname: Asadzadeh, Mohammad – sequence: 2 givenname: Suhail surname: Ahmad fullname: Ahmad, Suhail – sequence: 3 givenname: Ferry surname: Hagen fullname: Hagen, Ferry – sequence: 4 givenname: Jacques F. surname: Meis fullname: Meis, Jacques F. – sequence: 5 givenname: Noura surname: Al-Sweih fullname: Al-Sweih, Noura – sequence: 6 givenname: Ziauddin surname: Khan fullname: Khan, Ziauddin
BackLink	https://www.ncbi.nlm.nih.gov/pubmed/26580965$$D View this record in MEDLINE/PubMed
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Copyright	COPYRIGHT 2015 Public Library of Science 2015 Asadzadeh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. 2015 Asadzadeh et al 2015 Asadzadeh et al
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Notes	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Conceived and designed the experiments: MA SA NA ZK. Performed the experiments: MA FH. Analyzed the data: MA SA FH JFM NA ZK. Contributed reagents/materials/analysis tools: JFM ZK. Wrote the paper: MA SA FH JFM NA ZK. Competing Interests: The authors have declared that no competing interests exist.
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Snippet	Candida parapsilosis has now emerged as the second or third most important cause of healthcare-associated Candida infections. Molecular studies have shown that...
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SubjectTerms	Amplified fragment length polymorphism Analysis Antifungal Agents - pharmacology Candida Candida - classification Candida - drug effects Candida - genetics Candida - isolation & purification Candida albicans Candida parapsilosis Candidiasis Candidiasis - diagnosis Candidiasis - microbiology Care and treatment Chemical fingerprinting Clinical isolates Deoxyribonucleic acid Desorption Development and progression DNA DNA Fingerprinting - methods DNA sequencing DNA, Fungal - genetics DNA, Intergenic - genetics DNA, Ribosomal - genetics Economic aspects Epidemiology Fingerprinting Gene polymorphism Gene sequencing Genetic aspects Genomes Genotype Genotypes Haplotypes Health aspects Health care Hospitals Humans Identification Infections Infectious diseases Kuwait Lasers Low cost Mass spectrometry Medicine Microbial Sensitivity Tests Multiplex Polymerase Chain Reaction - economics Multiplex Polymerase Chain Reaction - methods Multiplexing Mycological Typing Techniques Pathogens Patient outcomes Patients Phylogeny Polymerase chain reaction Polymorphism Polymorphism, Restriction Fragment Length Scientific imaging Sequence Analysis, DNA Spacer Species Studies Yeast
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Title	Simple, Low-Cost Detection of Candida parapsilosis Complex Isolates and Molecular Fingerprinting of Candida orthopsilosis Strains in Kuwait by ITS Region Sequencing and Amplified Fragment Length Polymorphism Analysis
URI	https://www.ncbi.nlm.nih.gov/pubmed/26580965 https://www.proquest.com/docview/1734285157 https://www.proquest.com/docview/1735904636 https://pubmed.ncbi.nlm.nih.gov/PMC4651534 https://doaj.org/article/9687d9575c284438828e81c1621b57eb http://dx.doi.org/10.1371/journal.pone.0142880
Volume	10
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