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Sequences of primers used in Real-time PCR reaction in C_ albicans and C_ glabrata
| Gene | Primer | Sequence (5′->3′) |
|---|---|---|
| Sap1 | Forward | TGGGTTCCTGATGCTTCTGTT |
| Reverse | TCGGCAAAGACTTGCTTTGTG | |
| Sap2 | Forward | GGGGACATATGATCCAAGTGGT |
| Reverse | CCACCGGCTTCATTGGTTTT | |
| Sap3 | Forward | ATGTTACTGGTCCCCAAGGTG |
| Reverse | CCTTGACCAGCTTGACATGAA | |
| HWP1 | Forward | AATCATCAGCTCCTGCCACTG |
| Reverse | GTCGTAGAGACGACAGCACTA | |
| CDR1 | Forward | GGTGCTAATATCCAATGTTGG |
| Reverse | GTAATGGTTCTCTTTCAGCTG | |
| EPA1 | Forward | GGTCACTTACCCGCAAGCTA |
| Reverse | CCAGATGGCGTAGGCTTGAT | |
| ERG11 | Forward | GAGATTGCACCACCCATTGC |
| Reverse | TGGAGATAGCACCGAAACCG | |
| β-actin | Forward | ACGGTATTGTTTCCAACTGGGACG |
| Reverse | TGGAGCTTCGGTCAACAAAACTGG |
MIC result after CO2 exposure and MTBE in C_ albicans and C_ glabrata
| No | Spp | After 2 weeks with %5 CO2 | After 4 weeks with %5 CO2 | After 2 weeks with 5mg/dl MTBE | After 4 weeks with 5mg/dl MTBE | ||||
|---|---|---|---|---|---|---|---|---|---|
| Flu | Itra | Flu | Itra | Flu | Itra | Flu | Itra | ||
| TMML1 | C. glabrata | 4 | 1 | 4 | 1 | 2 | 2 | 32 | 8 |
| TMML2 | C. glabrata | 2 | 0.5 | 2 | 0.5 | 1 | 0.5 | 16 | 2 |
| TMML3 | C. glabrata | 2 | 0.5 | 4 | 0.5 | 0.5 | 1 | 16 | 2 |
| TMML4 | C. glabrata | 2 | 0.5 | 4 | 1 | 0.5 | 0.25 | 16 | 4 |
| TMML5 | C. glabrata | 2 | 0.5 | 2 | 0.5 | 2 | 0.5 | 16 | 4 |
| TMML6 | C. glabrata | 4 | 0.5 | 64 | 16 | 0.5 | 1 | 32 | 2 |
| TMML7 | C. glabrata | 2 | 0.5 | 2 | 0.5 | 0.5 | 0.062 | 0.5 | 0.125 |
| TMML8 | C. glabrata | 2 | 0.5 | 4 | 0.5 | 1 | 0.5 | 32 | 4 |
| TMML9 | C. glabrata | 4 | 1 | 64 | 16 | 1 | 1 | 16 | 8 |
| TMML10 | C. glabrata | 2 | 0.125 | 64 | 16 | 0.5 | 0.25 | 32 | 2 |
| TMML11 | C. albicans | 0.125 | 0.062 | 0.5 | 0.125 | 1 | 0.4 | 16 | 4 |
| TMML12 | C. albicans | 64 | 16 | 64 | 16 | 2 | 2 | 64 | 16 |
| TMML13 | C. albicans | 64 | 16 | 64 | 16 | 0.125 | 0.125 | 0.125 | 0.25 |
| TMML14 | C. albicans | 64 | 16 | 64 | 16 | 0.5 | 0.25 | 1 | 0.25 |
| TMML15 | C. albicans | 64 | 16 | 64 | 16 | 1 | 0.25 | 16 | 8 |
| TMML16 | C. albicans | 64 | 16 | 64 | 16 | 4 | 2 | 64 | 16 |
| TMML17 | C. albicans | 64 | 16 | 64 | 16 | 2 | 2 | 64 | 16 |
| TMML18 | C. albicans | 0.025 | 0.025 | 64 | 16 | 1 | 0.25 | 16 | 16 |
| TMML19 | C. albicans | 64 | 16 | 64 | 16 | 1 | 0.25 | 2 | 0.5 |
| TMML20 | C. albicans | 64 | 16 | 64 | 16 | 0.5 | 0.5 | 1 | 0.5 |
| TMML21 | C. albicans | 64 | 16 | 64 | 16 | 0.5 | 0.5 | 1 | 0.5 |
| TMML22 | C. albicans | 64 | 16 | 64 | 16 | 0.25 | 0.25 | 1 | 0.5 |
| TMML23 | C. albicans | 64 | 16 | 64 | 16 | 2 | 0.125 | 64 | 16 |
| TMML24 | C. albicans | 64 | 16 | 64 | 16 | 0.5 | 1 | 0.5 | 0.5 |
| TMML25 | C. albicans | 1 | 0.5 | 2 | 0.5 | 0.25 | 0.125 | 2 | 0.5 |
| TMML26 | C. albicans | 0.062 | 0.031 | 64 | 16 | 0.25 | 0.125 | 0.25 | 0.125 |
| TMML27 | C. albicans | 1 | 0.25 | 2 | 1 | 0.25 | 0.5 | 1 | 0.5 |
| TMML28 | C. albicans | 64 | 16 | 64 | 16 | 0.25 | 0.062 | 0.25 | 0.25 |
| TMML29 | C. albicans | 64 | 16 | 64 | 16 | 0.5 | 1 | 16 | 4 |
| TMML30 | C. albicans | 64 | 16 | 64 | 16 | 0.25 | 0.062 | 0.25 | 0.5 |
Isolates of C_ albicans and C_ glabrata which were sensitive for both drugs with PCR-RFLP
| TMML no | Source sample | MIC (µg/ml) Itr | MIC (µg/ml) Flu | Candida spp |
|---|---|---|---|---|
| TMML1 | BAL | 0.016 | 0.125 | albicans |
| TMML2 | sputum | 0.25 | 2 | albicans |
| TMML3 | sputum | 0.125 | 1 | albicans |
| TMML4 | sputum | 0.5 | 1 | albicans |
| TMML5 | mouth | 0.016 | 1 | albicans |
| TMML6 | nail | 0.5 | 1 | albicans |
| TMML7 | nail | 1 | 2 | albicans |
| TMML8 | BAL | 0.016 | 1 | albicans |
| TMML9 | nail | 0.062 | 0.5 | albicans |
| TMML10 | groin | 0.016 | 0.5 | albicans |
| TMML11 | BAL | 0.016 | 0.5 | albicans |
| TMML12 | BAL | 0.062 | 0.5 | albicans |
| TMML13 | sputum | 0.125 | 0.25 | albicans |
| TMML14 | sputum | 0.125 | 0.25 | albicans |
| TMML15 | sputum | 0.016 | 0.5 | albicans |
| TMML16 | sputum | 0.016 | 0.25 | albicans |
| TMML17 | skin | 0.5 | 2 | albicans |
| TMML18 | sputum | 0.125 | 0.5 | albicans |
| TMML19 | groin | 0.016 | 1 | albicans |
| TMML20 | sputum | 0.031 | 0.5 | albicans |
| TMML21 | stool | 1 | 8 | glabrata |
| TMML22 | sputum | 0.5 | 8 | glabrata |
| TMML23 | sputum | 1 | 16 | glabrata |
| TMML24 | sputum | 1 | 8 | glabrata |
| TMML25 | sputum | 0.25 | 8 | glabrata |
| TMML26 | sputum | 1 | 16 | glabrata |
| TMML27 | sputum | 0.125 | 16 | glabrata |
| TMML28 | sputum | 1 | 8 | glabrata |
| TMML29 | skin | 1 | 8 | glabrata |
| TMML30 | BAL | 0.062 | 1 | glabrata |
Dispersion the sources of Candida isolates
| Nature of specimen | C. albicans | C. glabrata | Other spices | Number of Candida spp | % |
|---|---|---|---|---|---|
| BAL | 8 | 3 | 8 | 19 | 18.02 |
| SPUTUM | 13 | 10 | 31 | 54 | 51.42 |
| NAIL | 6 | 2 | 4 | 12 | 11.42 |
| MOUTH | 2 | - | 2 | 4 | 3.8 |
| GROIN | 3 | - | 4 | 7 | 6.68 |
| SKIN | 1 | 1 | 4 | 6 | 5.81 |
| STOOL | - | 1 | 2 | 3 | 2.85 |
| Total | 33 | 17 | 60 | 105 |
Expression of CDR1, HWP1, SAP1-3 genes in comparison with β-actin in C_ albicans
| Gene | Type | Reaction Efficiency | Expression | Std. Error | 95% C.I. | P(H1) | Result |
|---|---|---|---|---|---|---|---|
| CDR1 | TRG | 1.0 | 1.623 | 0.160 - 20.966 | 0.035 - 68.781 | 0.342 | |
| HWP1 | TRG | 1.0 | 1.300 | 0.069 - 21.856 | 0.009 - 533.742 | 0.702 | |
| SAp3 | TRG | 1.0 | 2.298 | 0.471 - 20.190 | 0.077 - 48.176 | 0.042 | UP |
| Ssp2 | TRG | 1.0 | 4.547 | 0.388 - 86.223 | 0.007 - 849.223 | 0.024 | UP |
| Sap1 | TRG | 1.0 | 2.243 | 0.025 - 763.031 | 0.000 - 12,429.932 | 0.491 | |
| B-act | REF | 1.0 | 1.000 |
Expression of ERG11, EPA1, SAP3 genes in comparison with β-actin in C_ glabrata
| Gene | Type | Reaction Efficiency | Expression | Std. Error | 95% C.I. | P(H1) | Result |
|---|---|---|---|---|---|---|---|
| ERG11 | TRG | 1.0 | 0.876 | 0.045 - 29.445 | 0.000 - 123.640 | 0.894 | |
| EPA1 | TRG | 1.0 | 64.669 | 1.893 - 866.949 | 0.438 - 8,060.454 | 0.000 | UP |
| SAP3 | TRG | 1.0 | 0.745 | 0.009 - 17.387 | 0.000 - 434.218 | 0.784 | |
| B-act | REF | 1.0 | 1.000 |