Molecular heterogeneity of fluconazole‐resistant and ‐susceptible oral <i>Candida albicans</i> isolates within a single geographic locale

Dassanayake, R.S.; Ellepola, A. N. B.; Samaranayake, Y. H.; Samaranayak, L. P.

doi:10.1034/j.1600-0463.2002.100406.x

Cited by 16 publications

(11 citation statements)

References 34 publications

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“…Our result also showed that the DNA type of each isolate was patient specific and associated with ABC type and decade of isolation, but not associated with anatomical source of isolation, hospital origin, or fluconazole resistance patterns, which is in accordance with previous reports (8).…”

Section: Discussionsupporting

confidence: 81%

Multilocus Sequence Typing for Analyses of Clonality of Candida albicans Strains in Taiwan

Chen

et al. 2006

J Clin Microbiol

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Multilocus sequence typing (MLST) was used to characterize the genetic profiles of 51 Candida albicans isolates collected from 12 hospitals in Taiwan. Among the 51 isolates, 16 were epidemiologically unrelated, 28 were isolates from 11 critically ill, human immunodeficiency virus (HIV)-negative patients, and 7 were long-term serial isolates from 3 HIV-positive patients. Internal regions of seven housekeeping genes were sequenced. A total of 83 polymorphic nucleotide sites were identified. Ten to 20 different genotypes were observed at the different loci, resulting, when combined, in 45 unique genotype combinations or diploid sequence types (DSTs). Thirty (36.1%) of the 83 individual changes were synonymous and 53 (63.9%) were nonsynonymous. Due to the diploid nature of C. albicans, MLST was more discriminatory than the pulsed-field gel electrophoresis-BssHIIrestricted fragment method in discriminating epidemiologically related strains. MLST is able to trace the microevolution over time of C. albicans isolates in the same patient. All but one of the DSTs of our Taiwanese strain collections were novel to the internet C. albicans DST database (http://test1.mlst.net/). The DSTs of C. albicans in Taiwan were analyzed together with those of the reference strains and of the strains from the United Kingdom and United States by unweighted-pair group method using average linkages and minimum spanning tree. Our result showed that the DNA type of each isolate was patient specific and associated with ABC type and decade of isolation but not associated with mating type, anatomical source of isolation, hospital origin, or fluconazole resistance patterns.

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Section: Discussionsupporting

confidence: 81%

Multilocus Sequence Typing for Analyses of Clonality of Candida albicans Strains in Taiwan

Chen

et al. 2006

J Clin Microbiol

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“…Genotypic characterization in groups has been defined by Xu et al 28 for resistant isolates to fluconazole, who observed that resistant strains to this drug were genetically more similar to each other than they were to susceptible strains. Dassanayake et al 18 showed that, of the 20 isolates studied, only one resistant isolate presented the same profile as the susceptible one.…”

Section: Discussionmentioning

confidence: 99%

“…Previous works show that alterations in the ERG11 gene, responsible for ergosterol production, limit hyphal formation presumably because of the lack of ergosterol 27 . Although fluconazole is the most used antifungal agent for the treatment of infections caused by C. albicans, an increased number of resistant isolates have been observed 18 . Therefore, detection of resistance among isolates is considered of great importance.…”

Section: Discussionmentioning

confidence: 99%

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Molecular analysis and dimorphism of azole-susceptible and resistant Candida albicans isolates

Costa

Souza

Ataídes

et al. 2011

Rev. Soc. Bras. Med. Trop.

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Introduction: Candida albicans is responsible for superficial or systemic infections known as candidiasis, which may be found in infected tissue as unicellular budding yeasts, hyphae, or pseudohyphae. In this study, the effects of both fluconazole and itraconazole antifungal agents on the hyphal formation and genotypic characterization of C. albicans isolates classified as either susceptible or resistant were investigated. Methods: The hyphal production of five C. albicans isolates under the action of antifungal agents was investigated by culturing yeast on growth medium and on hyphal induction medium. The genotypic characterization was carried out for 13 isolates of C. albicans using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method. Results: The dimorphism analysis showed that the hyphal formation was higher in resistant than in the susceptible isolates to both azoles. The RAPD-PCR method identified the formation of two different groups. In group A, four resistant and two susceptible isolates were clustered, and in group B, one resistant and six susceptible isolates were clustered. Conclusions: Considering that hyphal formation was higher in resistant isolates in the presence of azole drugs, we confirmed that the hyphal production is closely related to susceptibility to azoles. These drugs may affect the morphogenesis of C. albicans depending on their susceptibility to these drugs. In relation to RAPD-PCR, most resistant isolates classified in group A and susceptible isolates in group B demonstrated that this method presented a similar standard between the two groups, suggesting that by this technique, a strong correlation between genotypes and fluconazole-resistant samples may be found.

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“…For RAPD, a single short primer of arbitrary sequence Ca2 (5'-GCGATCCCCA-3') was used to bind randomly to the target DNA resulting in the amplification of fragments of unknown sequence (15). The amplification reaction was carried out under the following conditions: 94°C for 5 minutes, followed by 35 cycles of 94°C for 30 seconds, 37°C for 30 seconds, 72°C for 90 seconds, and final extension at 72°C for 10 minutes.…”

Section: Molecular Analysismentioning

confidence: 99%

Characterization of the Same Mutations in FCA1 Gene Associated With 5-FC Resistance of Candida albicans

Deng

Wang

Ying

et al. 2017

Jundishapur J Microbiol

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Background: Infections with 5-Fluorocytosine (5-FC)-resistant Candida albicans isolates in China have rarely been reported in clinical settings. Here, we present two 5-FC-resistant C. albicans (Ca5508 and CaBD4291) strains, separately isolated from two patients in China. Objectives:The main aim of this study was to characterize the key genetic mutations responsible for 5-FC resistance of the two drug-resistant isolates from the first affiliated hospital of Nanchang university. Methods: Multiplex PCR and phenotypic tests were used to confirm the isolates Ca5508 and CaBD4291. The standard micro broth dilution method M27-A3 was used for susceptibility testing of the two isolates. Molecular typing and analysis were performed by the combination of random amplified polymorphic DNA (RAPD), multilocus sequence typing (MLST), amplification and Sanger sequencing of 5-FC resistance-associated genes (FUR1 and FCA1). Results: The two isolates Ca5508 and CaBD4291 were identified as C. albicans by multiplex-PCR and phenotypic tests. The antifungal susceptibility testing showed that both of the isolates were resistant to 5-FC (MIC > 64µg/mL); Ca5508 was also resistant to fluconazole, ketoconazole, and itraconazole whereas CaBD4291 was not. The RAPD analysis demonstrated that the two isolates belonged to the same genotype. MLST typing identified two different sequence types: isolate CaBD4191 for ST732 and isolate Ca5508 for ST2975 (a new ST). Although the two STs were generally related, ST732 differed from ST2975 with 3/7 loci (AAC1, SYA1, VPS13). Surprisingly, the two isolates represented completely identical sequences of the two drug-associated genes FUR1 and FCA1. Compared to the susceptible reference strain SC5314, Ca5508 and CaBD4291 were found to have no mutation in FUR1; however, three missense mutations at positions 31, 83, and 107 of FCA1 were identified. Conclusions: Here, we newly reported two 5-FC-resistant clinical isolates of C. albicans that represented the same three mutations in FCA1 gene contributed to 5-FC resistance of C. albicans.

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Molecular heterogeneity of fluconazole‐resistant and ‐susceptible oral Candida albicans isolates within a single geographic locale

Cited by 16 publications

References 34 publications

Multilocus Sequence Typing for Analyses of Clonality of Candida albicans Strains in Taiwan

Multilocus Sequence Typing for Analyses of Clonality of Candida albicans Strains in Taiwan

Molecular analysis and dimorphism of azole-susceptible and resistant Candida albicans isolates

Characterization of the Same Mutations in FCA1 Gene Associated With 5-FC Resistance of Candida albicans

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