About: Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing

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About: Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing
Creator	Wang, Bin Ellis, David Kong, Fanrong Pantarat, Namfon Wang, Huiping Widmer, Fred Xiao, Meng Sorrell, Tania Chen, Sharon Mcnicholas, Paul
Source	Medline; PMC
abstract	BACKGROUND: Amino acid substitutions in the target enzyme Erg11p of azole antifungals contribute to clinically-relevant azole resistance in Candida albicans. A simple molecular method for rapid detection of ERG11 gene mutations would be an advantage as a screening tool to identify potentially-resistant strains and to track their movement. To complement DNA sequencing, we developed a padlock probe and rolling circle amplification (RCA)-based method to detect a series of mutations in the C. albicans ERG11 gene using %22reference%22 azole-resistant isolates with known mutations. The method was then used to estimate the frequency of ERG11 mutations and their type in 25 Australian clinical C. albicans isolates with reduced susceptibility to fluconazole and in 23 fluconazole-susceptible isolates. RCA results were compared DNA sequencing. RESULTS: The RCA assay correctly identified all ERG11 mutations in eight %22reference%22 C. albicans isolates. When applied to 48 test strains, the RCA method showed 100% agreement with DNA sequencing where an ERG11 mutation-specific probe was used. Of 20 different missense mutations detected by sequencing in 24 of 25 (96%) isolates with reduced fluconazole susceptibility, 16 were detected by RCA. Five missense mutations were detected by both methods in 18 of 23 (78%) fluconazole-susceptible strains. DNA sequencing revealed that mutations in non-susceptible isolates were all due to homozygous nucleotide changes. With the exception of the mutations leading to amino acid substitution E266D, those in fluconazole-susceptible strains were heterozygous. Amino acid substitutions common to both sets of isolates were D116E, E266D, K128T, V437I and V488I. Substitutions unique to isolates with reduced fluconazole susceptibility were G464 S (n = 4 isolates), G448E (n = 3), G307S (n = 3), K143R (n = 3) and Y123H, S405F and R467K (each n = 1). DNA sequencing revealed a novel substitution, G450V, in one isolate. CONCLUSION: The sensitive RCA assay described here is a simple, robust and rapid (2 h) method for the detection of ERG11 polymorphisms. It showed excellent concordance with ERG11 sequencing and is a potentially valuable tool to track the emergence and spread of azole-resistant C. albicans and to study the epidemiology of ERG11 mutations. The RCA method is applicable to the study of azole resistance in other fungi.
has issue date	2009-08-14(xsd:dateTime)
bibo:doi	10.1186/1471-2180-9-167
bibo:pmid	19682357
has license	cc-by
sha1sum (hex)	2cbb038739adbd4487fa3c529e404189637e59d4
schema:url	https://doi.org/10.1186/1471-2180-9-167
resource representing a document's title	Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing
has PubMed Central identifier	PMC2782262
has PubMed identifier	19682357
schema:publication	BMC Microbiol
resource representing a document's body	covid:2cbb038739adbd4487fa3c529e404189637e59d4#body_text
is schema:about of	named entity 'substitutions' named entity 'rapid' named entity 'reduced' named entity 'fluconazole' named entity 'azole' named entity 'mutations' named entity 'frequency' named entity 'rolling circle amplification' named entity 'DNA sequencing' named entity 'isolates' named entity 'ROLLING CIRCLE' named entity 'BMC' named entity 'SERIES' named entity 'SCREENING' named entity 'MOLECULAR METHOD' named entity 'RESISTANT' named entity 'GENE' named entity 'RESISTANCE' named entity 'CLINICAL' named entity 'PROBE' named entity 'KNOWN' named entity 'RCA' named entity 'USING' named entity 'CANDIDA ALBICANS' named entity 'DNA SEQUENCING' named entity 'COMPARED' named entity 'TO TRACK' named entity 'ISOLATES' named entity 'SUSCEPTIBILITY' named entity 'TARGET' named entity 'ENZYME' named entity 'BASED' named entity 'FLUCONAZOLE' named entity 'TO DETECT' named entity 'SIMPLE' named entity 'TO IDENTIFY' named entity 'AMINO ACID SUBSTITUTIONS' named entity 'GENE MUTATIONS' named entity 'ROLLING CIRCLE' named entity 'CONTRIBUTE ' named entity 'THEIR' named entity 'MUTATIONS' named entity 'AMPLIFICATION' named entity 'RESULTS' named entity 'SUSCEPTIBLE' named entity 'ADVANTAGE' named entity 'CLINICAL' named entity 'AMPLIFICATION' named entity 'DETECTION' named entity 'ANTIFUNGALS' named entity 'ERG11' named entity 'METHOD' named entity 'ERG11' named entity 'RAPID' named entity 'ISOLATES' named entity 'SUSCEPTIBILITY' named entity 'REDUCED' named entity 'DNA SEQUENCING' named entity 'MOVEMENT' named entity 'REDUCED' named entity 'STRAINS' named entity 'CANDIDA ALBICANS' named entity 'DETECTION' named entity 'ESTIMATE' named entity 'GENE MUTATIONS' named entity 'FLUCONAZOLE' named entity 'FREQUENCY' named entity 'AUSTRALIAN' named entity 'TYPE' named entity 'AZOLE' named entity 'RAPID' named entity 'USED'

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