Sequential Induction of Drug Resistance and Characterization of an Initial Candida albicans Drug-Sensitive Isolate
Abstract
:1. Introduction
2. Materials and Methods
2.1. Candida albicans Isolates
2.2. DNA Extraction
2.3. DNA Sequencing and Sequence Analysis
2.3.1. Single-Nucleotide Polymorphism Detection
2.3.2. Annotation
2.4. Quantification of Ergosterol Content in the Plasma Membrane
2.5. Virulence Assay
2.6. Biofilm Formation Assay
2.7. Quantification of Cell Wall Chitin
2.8. Growth Kinetics
2.9. Statistical Analysis
3. Results
3.1. Ergosterol Quantification Assay
3.2. Chitin Quantification Assay
3.3. Biofilm Quantification Assay
3.4. Growth Rate Assay
3.5. Virulence Assay
3.6. WGS Analysis
3.6.1. Fluconazole Isolates
3.6.2. Caspofungin Isolates
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Isolate | Fluconazole Concentration |
---|---|
FL–MIC | 0.190 µg/mL |
FL–Int | 4.095 µg/mL |
FL–R | 8.000 µg/mL |
FL–2R | 16.000 µg/mL |
Isolate | Caspofungin Concentration |
---|---|
CS–MIC | 0.190 µg/mL |
CS–Int | 0.595 µg/mL |
CS–R | 1.000 µg/mL |
CS–2R | 2.000 µg/mL |
FL-MIC | FL-I | FL-R | FL-2R | ||
---|---|---|---|---|---|
Function | Genes | AA Change | AA Change | AA Change | AA Change |
Drug Resistance | ERG11 | L220L | L220L–L480L | L220L–L480L | L220L–T128K–L264S |
CDR2 | I704M–R365Q | I704M–M617L | I704M–M617L | R365Q–I704M–E374K | |
MSH2 | D73D | T141T | V127 | V127A | |
CHS3 | - | R762P | L321P–D388G–R762P | L321P–D388G–R762P | |
ERG4 | - | - | - | M157V | |
Stress response | SOD5 | - | A210P | A210P | A210P–G214D |
SOD6 | - | S225P | S225P | S225P | |
HSP60 | K67K | N281N | S267A | S267A | |
MNL1 | H349R | H349R–I175T | H349R–I175T | H349R–I175T | |
Phenotypic Switching/dimorphism | SAP1 | L53 | L53I | L53I | L53I |
CPH1 | T510A | T510A | T510A | T510A | |
EFG1 | K251K | K251K | I384T | I384T | |
MFG1 | - | A68P | A68P–Q76_Q80del | A68P–Q76_Q80del | |
SIR2 | L213 | L213 | L213 | L213S | |
RFG1 | - | - | Q118del | Q118del | |
FGR15 | V131A | V131A | V131A–S361L | V131A–S361L | |
Biofilm Formation and Immune Evasion | HWP1 | - | - | T296A–S441P | T296A–S441P |
ALS4 | I79V–V83I | I79V–V83I–V111A | I79V–V83I–V111A– S186P–D198N | I79V–V83I–V111A– S186P–D198N | |
NAG3 | - | R158* | Y278H | Y278H | |
UME6 | A146A | N257S–M267I | N257S–M267I | N257S–M267I –N396S | |
SFL1 | V691I | V691I | V691I–S373L | V691I–S373L |
CS-MIC | CS-I | CS-R | CS-2R | ||
---|---|---|---|---|---|
Function | Genes | AA Change | AA Change | AA Change | AA Change |
Drug Resistance | CDR2 | I704M | I704M–M617L | I704M–M617L–E374K | I704M–M617L–E374K–R365Q |
CRZ2 | - | - | S330A | S330A | |
GSC2 | A455A | A462P | A462P | A462P | |
Stress response | SOD5 | - | A210P | A210P–G214D | A210P–G214D |
SOD6 | - | S225P | S225P | S225P | |
HSP60 | S267A | S267A | S267A | S267A | |
MNL1 | H349R | H349R–I175T | H349R–I175T | R269 * | |
ATG26 | - | Y401C | E269K–Y401C | E269K–Y401C–T414I | |
Phenotypic Switching/dimorphism | CPH1 | P451P | I79I | T510A | T510A |
MFG1 | - | - | Q80del | Q80del | |
RFG1 | - | - | T39S | T39S | |
SIR2 | - | - | L213S | L213S | |
FGR15 | - | S361L | S361L | S361L | |
Biofilm Formation and Immune Evasion | HWP1 | - | S441P | S441P–T296A | S441P–T296A |
ALS4 | - | I79V–V83I–V111A | I79V–V83I–V111A–S186P | I79V–V83I–V111A–S186P–D198N | |
NAG3 | - | - | Y278H | R158 * | |
UME6 | N257S–M267I | N257S–M267I | N257S–M267I–N396S | N257S–M267I–N396S–N484del | |
SFL1 | V691I–S373L | V691I–S373L | V691I–S373L–T75A | T75A–Q301fs |
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El Hachem, S.; Fattouh, N.; Chedraoui, C.; Finianos, M.; Bitar, I.; Khalaf, R.A. Sequential Induction of Drug Resistance and Characterization of an Initial Candida albicans Drug-Sensitive Isolate. J. Fungi 2024, 10, 347. https://doi.org/10.3390/jof10050347
El Hachem S, Fattouh N, Chedraoui C, Finianos M, Bitar I, Khalaf RA. Sequential Induction of Drug Resistance and Characterization of an Initial Candida albicans Drug-Sensitive Isolate. Journal of Fungi. 2024; 10(5):347. https://doi.org/10.3390/jof10050347
Chicago/Turabian StyleEl Hachem, Setrida, Nour Fattouh, Christy Chedraoui, Marc Finianos, Ibrahim Bitar, and Roy A. Khalaf. 2024. "Sequential Induction of Drug Resistance and Characterization of an Initial Candida albicans Drug-Sensitive Isolate" Journal of Fungi 10, no. 5: 347. https://doi.org/10.3390/jof10050347