Table 5. Characteristics of the included studies using molecular methods for evaluating fungal presence in teeth with secondary endodontic infection.
| Study Country | No. of subjects and characteristics | No. of samples and characteristics | Procedures before collection | Sample collection | Yeast identification method | Yeast spp. recovered (number of cases; prevalence) | Risk of bias |
|---|---|---|---|---|---|---|---|
| Siqueira & Rôças 2004 [55] Brazil | 22 Age range 29–80 yrs, mean age 46 yrs |
22 RCT completed > 2 y earlier, no direct exposure to oral cavity, no periodontal pocket |
Pumice cleansing, rubber dam isolation, 3% H2O2, 2.5% NaOCl. Endodontic access, 2.5% NaOCl, mechanical removal of root filling (no solvent). Introduction of sterile saline into the canals | (a) #15 K-type file (b) 3 paper points; placed in TE buffer and frozen at -20°C |
Primer for C. albicans toward act1 gene for actin: GCC GGT GAC GAC GCT CCA AGA GCT G CCG TGT TCA ATT GGG TAT CTC AAG GTC Positive control strains: C. albicans ATCC 10231, ATCC 44858 |
C. albicans (2; 9.09%) | 0 |
| Rôças et al. 2008 [63] Germany | 13 Age range 22–60 yrs, mean age 43.5 yrs |
17 RCT completed at least 1 year earlier, no direct exposure to oral cavity |
Rubber dam isolation, 3% H2O2, 3% NaOCl. Endodontic access, 3% NaOCl. Mechanical removal of root filling (no solvent). Introduction of sterile saline into the canals if dried | 2 paper points; placed in TE buffer | Primer for C. albicans, toward act1 gene for actin: F: GCC GGT GAC GAC GCT CCA AGA GCT G R: CCG TGT TCA ATT GGG TAT CTC AAG GTC Positive control strains: C. albicans ATCC 10231, ATCC 44858 |
C. albicans (1; 5.88%) | 0 |
| Miranda et al. 2009 [64] Brazil | 168 No atb and antifungal treatment during the previous 6 months, no systemic diseases; 20-65yrs |
184 Periapical radiolucency, no pulp exposure (only one root sampled from multi-rooted, widest) |
Rubber dam isolation; endodontic access (no water spray), disinfection (30% H2O2 for 1 min, 5% iodine for 1 min, 5% Na2S2O3 for 1 min), sterility control sample; Introduction of sterile saline into the canals | 3 paper points for 1 min, placed in modified Sabouraud broth with 100 mg L-1 chloramphenicol | PCR fingerprint technique. Primer: EI1: CTG GCT TGG TGT ATG T Positive control strains: C. albicans ATCC 18804, C. parapsilosis ATCC 22019, C. krusei ATCC 2159, C. tropicalis UFMG- A10, C. glabrata NCYC 388, C. lusitaniae CBS 6936, C. dubliniensis CBS 7987. Different genetic fingerprint profile was identified by sequencing the D1/D2 variable domains of the large subunit rDNA (primers: NL-1, NL-4) |
Patient-level analysis: 16 teeth randomly excluded form analysis. C. albicans (34) C. parapsilosis (3), Total Candida spp. (37, 22.6%) |
0 |
| Anderson et al. 2012 [69] Germany | 21 no systemic diseases, no atb in the last 30 days |
20 (1 tooth out of 21 excluded for contamination of quality control sample) RCT completed at least 2 yrs earlier, no direct exposure to oral cavity, asymptomatic |
Rubber dam isolation, disinfection (30% H2O2, 2,5% NaOCl), endodontic access, disinfection (30% H2O2, 2,5% NaOCl, 5% Na2S2O3), sterility control samples (foam pellets), mechanical removal of root filling (no solvent). Introduction of sterile saline into the canals | 3 paper points for 1 min; placed in RTF, frozen at -20°C | 18S-rRNA genes amplification with primers: ITS1: CTT GGT CAT TTA GAG GAA GTA A ITS4: TCC TCC GCT TAT TGA TAT GC |
Fungal species (1; 5%) | 0 |
| Dumani et al. 2012 [71] Turkey | 170 (100 primary, 70 secondary) M: f = 63:107 |
231 (117 primary, 114 secondary) | Pumice cleansing, rubber dam isolation, disinfection (35% H2O2, 5% NaOCl), endodontic access, disinfection (5% NaOCl, Na2S2O3), sterility control samples (cotton pellets), eventually mechanical removal of root filling (no solvent), introduction of sterile saline into the canals | 3 paper points for 1 min; placed in TE buffer, frozen at -20°C | Primer for C. albicans toward act1 gene for actin: F: GCC GGT GAC GAC GCT CCA AGA GCT G R: CCG TGT TCA ATT GGG TAT CTC AAG GTC Positive control strains: C. albicans ATCC 90028 |
Primary: C. albicans (23; 19.66%) Secondary: C. albicans (13; 11.40%) |
0 |
| Poptani et al. 2012 [73] India | NS no antibiotic therapy in the last 1 month |
20 Symptomatic; RTC completed at least 1 yr. earlier, 1 root per tooth was sampled (the one associated to lesion and/or widest) |
Pumice cleansing, rubber dam isolation, disinfection (3% H2O2, 2.5% NaOCl), endodontic access (with saline irrigation), disinfection (2.5% NaOCl), mechanical removal of root filling (no solvent), introduction of sterile saline into the canals | 2–3 paper points for 1 min; placed in TE buffer and frozen at -20°C | Primer for C. albicans: f: GCC GGT GAC GAC GCT CCA AGA GCT G r: CCG TCA GGG GAC GTT CAG Positive control strains: C. albicans ATCC 10231 |
C. albicans (7; 35%) | 0 |
| Karygianni et al. 2015 [75] Germany | 5 No periodontitis, no severe systemic diseases, no antibiotic therapy in the last 30 days |
5 First lower molars, asymptomatic, RCT completed at least 2 yrs earlier, no direct exposure to oral cavity |
Disinfection (30% H2O2, 2.5% NaOCl), rubber dam isolation, endodontic access, disinfection (30% H2O2, 2.5% NaOCl, 5% Na2S2O3), sterility control sample, mechanical root-filling removal (no solvents), introduction of sterile saline into the canal | a) Obturation material samples; placed in RTF (b) 3 paper points for 1 min; placed in RTF and frozen at -80°C |
18S-rRNA genes amplification with primers: ITS1: CTT GGT CAT TTA GAG GAA GTA A ITS4: TCC TCC GCT TAT TGA TAT GC |
Fungal species 0 | 1 |
| Zargar et al. 2019 [81] Iran | 30 No severe systemic diseases, no antibiotic therapy in the last 30 days, age range 23–70 yrs |
30 | Disinfection (30% H2O2, 3% NaOCl), caries/restoration removal and endodontic access with sterile bur, rubber dam isolation, disinfection (30% H2O2, 3% NaOCl, 5% Na2S2O3), sterility control sample; mechanical root-filling removal (no solvents). introduction of sterile saline into the canal | (a) #20 and #25 Haedstrom file; placed in thioglycolate (b) #20 and #25 paper point for 1 min; placed in thioglycolate |
Primer: ITS3: GCA TCG ATG AAG AAC GCA GC |
C. albicans (10; 33.33%) | 0 |
Atb = antibiotics; ATCC = American Type Culture Collection (Rockville, MD); °C = degree Celsius; F = forward; f = females; H2O2 = hydrogen peroxide; M = males; min = minutes; Na2S2O3 = sodium thiosulfate; NaOCl = sodium hypochlorite; NS = not specified; PCR = polymerase chain reaction; R = reverse; RCT = root canal treatment; RTF = reduced transport fluid; spp. = species; T2DM = type 2 diabetes mellitus; TE = Tris–ethylenediaminetetraacetic acid; TSB-DMSO = trypticase-soy broth with 5% dimethyl sulfoxide; VMGA = Viability Medium Göteborg Agar; yrs = years. RISK OF BIAS: (0 low risk; 1–2 moderate risk; 3–5 high risk)