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Prevalence of Closely Related Candida albicans Species among Patients with Vulvovaginal Candidiasis in Southern Poland Based on the hwp1 Gene Amplification

KAROLINA KLESIEWICZ
KAROLINA KLESIEWICZ
, 
PAULINA MROWIEC
PAULINA MROWIEC
, 
KATARZYNA KANIA
KATARZYNA KANIA
, 
IWONA SKIBA-KUREK
IWONA SKIBA-KUREK
, 
JOANNA BIAŁECKA
JOANNA BIAŁECKA
, 
MAGDALENA NAMYSŁ
MAGDALENA NAMYSŁ
 et 
MARIANNA MAŁEK
MARIANNA MAŁEK
   | 24 mars 2023
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Article Category: ORIGINAL PAPER
Publié en ligne: 24 mars 2023
Pages: 69 - 77
Reçu: 21 déc. 2022
Accepté: 13 févr. 2023
DOI: https://doi.org/10.33073/pjm-2023-011
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© 2023 KAROLINA KLESIEWICZ et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Morphology of reference strains of yeasts colonies after 48 h incubation in 37°C on chromID® Candida (bioMérieux, France). A. a) Candida albicans ATCC® 90028™ (dark blue colonies), b) Candida glabrata ATCC® 2001™ (white colonies), c) Candida tropicalis ATCC® 9968™ (pink colonies). B. a) Candida africana ATCC® MYA 2669™ (small, light blue colonies), b) Candida albicans ATCC® 90028™ (dark blue colonies), c) Candida dubliniensis clinical strain (turquoise colonies).
Morphology of reference strains of yeasts colonies after 48 h incubation in 37°C on chromID® Candida (bioMérieux, France). A. a) Candida albicans ATCC® 90028™ (dark blue colonies), b) Candida glabrata ATCC® 2001™ (white colonies), c) Candida tropicalis ATCC® 9968™ (pink colonies). B. a) Candida africana ATCC® MYA 2669™ (small, light blue colonies), b) Candida albicans ATCC® 90028™ (dark blue colonies), c) Candida dubliniensis clinical strain (turquoise colonies).

Fig. 2.

Representative samples of the hwp1 gene PCR assay visualised in electrophoresis. Line 1 and 13 – molecular weight marker (O’GeneRuler 50 bp DNA Ladder; Thermo Scientific, USA) Line 2 – positive control of Candida albicans ATCC® 90028™ (product size 941 bp). Line 3 – positive control of Candida dubliniensis NCPF 3949 (product size 569 bp). Line 4 – positive control of Candida africana ATCC® MYA 2669™ (product size 700 bp) Line 5, 6, 7, 9, 11 – clinical Candida albicans strains. Line 8, 10 – clinical Candida dubliniensis strains. Line 12 – negative control
Representative samples of the hwp1 gene PCR assay visualised in electrophoresis. Line 1 and 13 – molecular weight marker (O’GeneRuler 50 bp DNA Ladder; Thermo Scientific, USA) Line 2 – positive control of Candida albicans ATCC® 90028™ (product size 941 bp). Line 3 – positive control of Candida dubliniensis NCPF 3949 (product size 569 bp). Line 4 – positive control of Candida africana ATCC® MYA 2669™ (product size 700 bp) Line 5, 6, 7, 9, 11 – clinical Candida albicans strains. Line 8, 10 – clinical Candida dubliniensis strains. Line 12 – negative control

Fig. 3.

Susceptibility of 309 Candida albicans clinical isolates caused VVC obtained by Fungitest™ (BioRad, France). S – susceptible, I – intermediate (referring to “susceptible, increased exposure” according to the EUCAST recommendations), R – resistant.
Susceptibility of 309 Candida albicans clinical isolates caused VVC obtained by Fungitest™ (BioRad, France). S – susceptible, I – intermediate (referring to “susceptible, increased exposure” according to the EUCAST recommendations), R – resistant.

Percentage of compatible results obtained by the methods used to discriminate species among Candida albicans complex (VITEK®2 system and MALDI-TOF MS) with the hwp1 gene amplification with PCR.

The compliance with PCR assayNo. (%) n = 326
VITEK 2 319 (97.85) 7 (2.15)
MALDI-TOF MS 323 (99.08) 3 (0.92)

Molecular discrimination among Candida albicans complex.

Species No. (%) n = 326
Candida albicans 307 (94.17)
Candida dubliniensis 17 (5.21)
Candida albicans + Candida dubliniensis 2 (0.61)

Discrepancies between species identification among Candida albicans complex obtained by the Vitek®2 system, MALDI-TOF MS and the hwp1 gene amplification with PCR.

Isolate No. VITEK 2 MALDI-TOF MS PCR assay based on the hwp1 gene amplification
Species identification Confidence level (probability) Species identification Confidence level Species identification Product size (bp)
CA67 clinical strain Candida albicans/Candida dubliniensis low (< 85%) Candida dubliniensis 99.90% Candida dubliniensis 569
CA95 clinical strain Candida albicans excellent (96%) Candida albicans 99.90% Candida albicans + Candida dubliniensis 941 + 569
CA98 clinical strain not identified – Candida dubliniensis 99.90% Candida dubliniensis 569
CA125 clinical strain Candida ciferrii acceptable (86%) Candida dubliniensis 99.90% Candida dubliniensis 569
CA13 clinical strain Candida albicans/Candida dubliniensis low (< 85%) Candida dubliniensis 99.90% Candida dubliniensis 569
CA137 clinical strain Candida albicans good (91%) Candida albicans 99.90% Candida dubliniensis 569
CA313 clinical strain Candida albicans very good (95%) Candida albicans 99.90% Candida albicans + Candida dubliniensis 941 + 569
Candida albicansATCC® 90028™ reference strain Candida albicans excellent (98%) Candida albicans 99.90% Candida albicans 941
Candida dubliniensisNCPF 3949 reference strain Candida dubliniensis very good (93%) Candida dubliniensis 99.90% Candida dubliniensis 569
Candida africanaMYA 2669™ reference strain Candida dubliniensis/Candida parapsilosis low (< 85%) Candida albicans 99.90% Candida africana 700

Prevalence of single VVC and co-occurrence with bacteria.

Type of co-occurrence No. (%)n = 326
Single VVC 197 (60.43)
VVC + bacteria: 129 (39.57)
  Gardnerella vaginalis   60 (18.40)
  Streptococcus agalactiae   41 (12.58)
  Gardnerella vaginalis + Streptococcus agalactiae    11 (3.37)
  Escherichia coli      9 (2.76)
  Escherichia coli + Streptococcus agalactiae      3 (0.92)
  Gardnerella vaginalis + Escherichia coli      2 (0.61)
  Methicillin susceptible Staphylococcus aureus      1 (0.31)
  Streptococcus pyogenes      1 (0.31)
  Klebsiella pneumoniae + Streptococcus agalactiae      1 (0.31)
eISSN:
2544-4646
Langue:
Anglais
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Sujets de la revue:
Life Sciences, Microbiology and Virology
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