Background
Methods
Subjects
Patient
a
| Subject group
b
| Collection site | Sex
c
| Age | Lesion severity
d
| Episodee
| Duration of disease [yrs] | Treatment
f
| Family history of disease | Other diseases
g
| Strain no. | Species identifiction
h
| |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1.
| TK | AD | Head | M | 24 | 37 | R | 24 | Topical; sys: antihistaminics | Positive | No | 40.10.I. |
M. sympodialis
|
2.
| Face | 40.10.II. |
M. sympodialis
| ||||||||||
3.
| Back | 40.10.IV. |
M. sympodialis
| ||||||||||
4.
| MP | Head | F | 28 | 31 | R | 23 | Topical; sys: antihistaminics | Positive | Rhinoconjunctivitis | 8.11.I. |
M. sympodialis
| |
5.
| Chest | 8.11.III. |
M. sympodialis + C. d.
*
| ||||||||||
6.
| Back | 8.11.IV. |
M. sympodialis
| ||||||||||
7.
| DJ | Chest | M | 29 | 34.6 | R | 29 | Topical; sys: antihistaminics | Positive | Rhinoconjunctivitis | 27.10.III. |
M. sympodialis
| |
8.
| Back | 27.10.IV. |
M. sympodialis
| ||||||||||
9.
| JW | Chest | F | 22 | 10.2 | R | 22 | Topical; sys: antihistaminics | Positive | No | 7.11.III. |
M. sympodialis
| |
10.
| Back | 7.11.IV. |
M. sympodialis
| ||||||||||
11.
| MB | Back | M | 22 | 39.4 | R | 22 | Topical; sys: antihistaminics | Negative | 25.10.IV. |
M. sympodialis
| ||
12.
| EP | Back | F | 31 | 10.2 | R | 31 | Topical; sys: antihistaminics | Negative | Vitiligo | 17.10.IV. |
M. sympodialis
| |
13.
| BT | PS | Head | F | 50 | 8.2 | R | 20 | Topical; sys: CyA | Positive | No | 10.11.I. |
M. sympodialis
|
14.
| Face | 10.11.II. |
M. sympodialis + A. p.
**
| ||||||||||
15.
| Chest | 10.11.IIIA. |
M. furfur
| ||||||||||
16.
| Back | 10.11.IV. |
M. sympodialis
| ||||||||||
17.
| BH | Face | F | 44 | 10 | R | 15 | Topical | Negative | No | 45.08.II. |
M. furfur
| |
18.
| Chest | 45.08.III. |
M. furfur + M. sympodialis
| ||||||||||
19.
| SK | Back | M | 38 | 3.6 | R | 5 | Topical; sys: CS | Negative | No | 20.09.IV. |
M. furfur + M. sympodialis
| |
20.
| EN | Back | F | 31 | 6.6 | R | 5 | Topical | Negative | No | 17.09.IV. |
M. sympodialis
| |
21.
| KK | Face | M | 23 | 11.2 | R | 12 | Topical; sys: CyA | Positive | No | 43.08.II. |
M. furfur
| |
22.
| MB | Back | F | 29 | 5.7 | R | 15 | Topical | Positive | No | 6.11.IV. |
M. sympodialis
| |
23.
| MT | Control | Head | F | 31 | No | 1.11.IA. |
M. globosa
| |||||
24.
| Face | 1.11.II. |
M. globosa + M. restricta
| ||||||||||
25.
| Chest | 1.11.IIIA. |
M. sympodialis
| ||||||||||
26.
| Back | 1.11.IVA. |
M. sympodialis
| ||||||||||
27.
| CM | Chest | M | 70 | No | 69.09.III. |
M. slooffiae
| ||||||
28.
| Back | 69.09.IV. |
M. slooffiae + M. sympodialis
| ||||||||||
29.
| BP | Chest | M | 69 | No | 67.09.III. |
M. sympodialis
| ||||||
30.
| Back | 67.09.IV. |
M. sympodialis
| ||||||||||
31.
| PK | Chest | M | 27 | No | 68.09.III. |
M. sympodialis
| ||||||
32.
| Back | 68.09.IV. |
M. sympodialis
| ||||||||||
33.
| MS | Chest | F | 28 | No | 2.11.III. |
M. sympodialis
| ||||||
34.
| Back | 2.11.IV. |
M. sympodialis
| ||||||||||
35.
| JP | Back | F | 31 | No | 12.09.IV. |
M. sympodialis
|
Sample collection
Culture conditions and yeast strains
Drug susceptibility testing
DNA isolation
Species identification
Identification by phenotypic methods
Molecular analysis
PCR-RFLP analysis
PCR-sequencing
Strain no. | Species identification by means of | Final species identification
d
| ||||||||
---|---|---|---|---|---|---|---|---|---|---|
Phenotypic methods
a
| Molecular methods | |||||||||
PCR-RFLP
b
| PCR sequencing
c
| |||||||||
ITS accession no. | Similarity with ITS of [%]: | D1/D2 accession no. | Similarity with D1/D2 of [%]: | |||||||
1.
| 40.10.I. |
M. slooffiae
|
M. sympodialis
| KC141968 |
M. sympodialis MA 73 | 100% | KC415092 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
2.
| 40.10.II. |
M. slooffiae
|
M. sympodialis
| KC141969 |
M. sympodialis CBS 7222 | 100% | KC415093 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
3.
| 40.10.IV. |
M. sympodialis
|
M. sympodialis
| KC141970 |
M. sympodialis CBS 7222 | 100% | KC415094 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
4.
| 8.11.I. |
M. obtusa
|
M. sympodialis
| KC152895 |
M. sympodialis CBS 7222 | 100% | KC415085 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
5.
| 8.11.III. |
M. furfur
|
M. sympodialis
| KC152904 |
C. diffluens CBS 160 | 100% | KC241877 |
C. diffluens CBS 6436 | 100% |
M. sympodialis + C. d.
*
|
6.
| 8.11.IV. |
M. sympodialis
|
M. sympodialis
| KC152896 |
M. sympodialis CBS 7222 | 100% | KC415086 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
7.
| 27.10.III. |
M. sympodialis
|
M. sympodialis
| KC141966 |
M. sympodialis MA 73 | 100% | KC415095 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
8.
| 27.10.IV. |
M. sympodialis
|
M. sympodialis
| KC141967 |
M. sympodialis MA 73 | 100% | KC415096 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
9.
| 7.11.III. |
M. sympodialis
|
M. sympodialis
| KC152893 |
M. sympodialis CBS 7222 | 100% | KC415083 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
10.
| 7.11.IV. |
M. sympodialis
|
M. sympodialis
| KC152894 |
M. sympodialis MA 73 | 100% | KC415084 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
11.
| 25.10.IV. |
M. sympodialis
|
M. sympodialis
| KC119577 |
M. sympodialis CBS 7222 | 100% | KC415097 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
12.
| 17.10.IV. |
M. sympodialis
|
M. sympodialis
| KC119576 |
M. sympodialis CBS 7222 | 100% | KC415098 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
13.
| 10.11.I. |
M. sympodialis
|
M. sympodialis
| KC152897 |
M. sympodialis CBS 7222 | 100% | KC415087 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
14.
| 10.11.II. |
M. sympodialis
|
M. sympodialis
| KC152905 |
A. pullulans CPC 13701 | 100% | KC241878 |
A. pullulans RA406 | 100% |
M. sympodialis + A. p.
**
|
15.
| 10.11.IIIA. |
M. furfur
|
M. furfur
| KC152897 |
M. furfur M235 | 100% | KC415088 |
M. furfur VG Ig 02 | 99% |
M. furfur
|
16.
| 10.11.IV. |
M. sympodialis
|
M. sympodialis
| KC152901 |
M. sympodialis CBS 7222 | 100% | KC415091 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
17.
| 45.08.II. |
M. furfur
|
M. furfur
| KC141972 |
M. furfur M235 | 100% | KC415099 |
M. furfur VG Ig 02 | 100% |
M. furfur
|
18.
| 45.08.III. |
M. furfur
|
M. furfur
| KC141973 |
M. sympodialis CBS 7222 | 100% | KC415100 |
M. sympodialis IFM 48109 | 99% |
M. furfur + M. sympodialis
|
19.
| 20.09.IV. |
M. furfur
|
M. furfur + M. sympodialis
| KC141965 |
M furfur M235 | 100% | KC415101 |
M. furfur VG Ig 02 | 99% |
M. furfur + M. sympodialis
|
20.
| 17.09.IV. |
M. sympodialis
|
M. sympodialis
| KC109788 |
M. sympodialis MA 73 | 100% | KC415102 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
21.
| 43.08.II. |
M. furfur
|
M. furfur
| KC141971 |
M. furfur M235 | 100% | KC415103 |
M. furfur VG Ig 02 | 100% |
M. furfur
|
22.
| 6.11.IV. |
M. sympodialis
|
M. sympodialis
| KC152892 |
M. sympodialis MA 73 | 100% | KC415082 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
23.
| 1.11.IA. |
M. globosa
|
M. globosa
| KC152884 |
M. globosa CBS 7966 | 96% | KC415074 |
M. sympodialis IFM 48109 | 100% |
M. globosa
|
24.
| 1.11.II. |
M. globosa
|
M. globosa + M. restricta
| KC152885 |
M. restricta MRE28 | 99% | KC415075 |
M. sympodialis IFM 48109 | 100% |
M. globosa + M. restricta
|
25.
| 1.11.IIIA. |
M. sympodialis
|
M. sympodialis
| KC152886 |
M. sympodialis MA 73 | 100% | KC415076 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
26.
| 1.11.IVA. |
M. furfur
|
M. sympodialis
| KC152888 |
M. sympodialis MA 73 | 100% | KC415078 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
27.
| 69.09.III. |
M. slooffiae
|
M. slooffiae
| KC141978 |
M. slooffiae CBS 7956 | 99% | KC415104 |
M. slooffiae sc7LG | 99% |
M. slooffiae
|
28.
| 69.09.IV. |
M. slooffiae
|
M. slooffiae
| KC141979 |
M. slooffiae CBS 7956 | 99% | KC415105 |
M. sympodialis IFM 48109 | 100% |
M. slooffiae + M. sympodialis
|
29.
| 67.09.III. |
M. sympodialis
|
M. sympodialis
| KC141974 |
M. sympodialis CBS 7222 | 100% | KC415106 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
30.
| 67.09.IV. |
M. slooffiae
|
M. sympodialis
| KC141975 |
M. sympodialis CBS 7222 | 100% | KC415107 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
31.
| 68.09.III. |
M. sympodialis
|
M. sympodialis
| KC141976 |
M. sympodialis CBS 7222 | 100% | KC415108 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
32.
| 68.09.IV. |
M. sympodialis
|
M. sympodialis
| KC141977 |
M. sympodialis CBS 7222 | 100% | KC415109 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
33.
| 2.11.III. |
M. sympodialis
|
M. sympodialis
| KC152890 |
M. sympodialis MA 73 | 100% | KC415080 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
34.
| 2.11.IV. |
M. obtusa
|
M. sympodialis
| KC152891 |
M. sympodialis MA 73 | 100% | KC415081 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
35.
| 12.09.IV. |
M. sympodialis
|
M. sympodialis
| JX915741 |
M. sympodialis MA 73 | 100% | KC415110 |
M. sympodialis IFM 48109 | 100% |
M. sympodialis
|
Statistical analysis
Ethics
Results
Phenotypic identification
PCR-RFLP analysis
PCR-sequence analysis
Comparison of PCR-RFLP analysis and PCR-sequencing analysis
Comparison of phenotypic and molecular methods
Drug susceptibility testing
Malassezia sp. (no.) | Antifungal drug
a
| |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
CPO | FLZ | KTZ | ITZ | ECZ | MNZ | |||||||
Zone [mm]
b
| Cat. (no.)
c
| Zone [mm] | Cat.(no.) | Zone [mm] | Cat. (no.) | Zone [mm] | Cat. (no.) | Zone [mm] | Cat. (no.) | Zone [mm] | Cat. (no.) | |
M. sympodialis (13) | 28 | S (13) | 43 | S (13) | 33.9 | S (13) | 36.5 | S (13) | 24.5 | S (12) | 26.5 | S (13) |
17 | I (1) | |||||||||||
M. furfur (2) | 23 | S (2) | 36 | S (2) | 30 | S (1) | 29 | S (2) | 15.5 | I (2) | 20 | S (1) |
25 | I (1) | |||||||||||
17 | I (1) | |||||||||||
M. furfur + M. sympodialis (2) | 22.5 | S (2) | 39 | S (2) | 25.5 | I (2) | 28.5 | S (2) | 14.5 | I (2) | 20 | S (1) |
15 | I (1) | |||||||||||
M. slooffiae (1) | 30 | S (1) | 45 | S (1) | 35 | S (1) | 45 | S (1) | 30 | S (1) | 20 | S (1) |
M. slooffiae + M. sympodialis (1) | 30 | S (1) | 40 | S (1) | 40 | S (1) | 40 | S (1) | 30 | S (1) | 16.5 | I (1) |