Genotypic analysis of secreted aspartyl proteinases in vaginal <i>Candida albicans</i> isolates

Lima, Janaína S.; Braga, Kaoane Regina G. S.; Vieira, Camila A.; Souza, Wérika Weryanne R.; Chávez-Pavoni, Juliana Helena; Araújo, Claudinéia de; Goulart, Letícia S.

doi:10.5935/1676-2444.20180006

LABORATORY MEDICINE, ORIGINAL ARTICLE • J. Bras. Patol. Med. Lab. 54 (1) • Jan-Feb 2018 • https://doi.org/10.5935/1676-2444.20180006 linkcopy

Genotypic analysis of secreted aspartyl proteinases in vaginal Candida albicans isolates

Análise genotípica de aspartil proteases secretórias em isolados vaginais de Candida albicans

Authorship SCIMAGO INSTITUTIONS RANKINGS

Introduction: Candida albicans is the most common etiologic agent of fungal vaginitis. These yeasts produce secreted aspartyl proteinases encoded by a family of 10 genes (SAP1-10).

Objective: The purpose of this study was to analyze the presence of genes SAP1-7 in vulvovaginal C. albicans.

Materials and method: The study included 26 C. albicans vaginal isolates. Detection of aspartyl proteases genes (SAP1-7) was performed by polymerase chain reaction (PCR).

Results: The most frequent gene in C. albicans isolated from colonization was SAP6 (93.33%), and from infection, SAP7 (100%). We observed a statistical difference (p = 0.049) in SAP1 gene frequency between isolates from vulvovaginal colonization and infection.

Conclusion: High frequency of SAP genes was observed in vulvovaginal C. albicans. The results suggest SAP1 participation in vulvovaginal candidiasis infection.

Key words:
vulvovaginal candidiasis; Candida albicans; virulence factors

Primer	Oligonucleotide sequence	Size (pb)
SAP 1	Forward 5'-TCA ATC AAT TTA CTC TTC CAT TTC TAA CA-3' Reverse 5'-CCA GTA GCA TTA ACA GGA GTT TTA ATG ACA-3'	161
SAP 2	Forward 5'-AAC AAC AAC CCA CTA GAC ATC ACC-3' Reverse 5'-TGA CCA TTA GTA ACT GGG AAT GCT TTA GGA-3'	178
SAP 3	Forward 5'-CCT TCT CTA AAA TTA TGG ATT GGA AC-3' Reverse 5'-TTG ATT TCA CCT TGG GGA CCA GTA ACA TTT-3'	231
SAP 4	Forward 5'-TTA TTT TTA GAT ATT GAG CCC ACA GAA A-3' Reverse 5'-GCC AGT GTC AAC AAT AAC GCT AAG TT-3'	171
SAP 5	Forward 5'-AGA ATT TCC CGT CGA TGA GAC TGG T-3' Reverse 5'-CAA ATT TTG GGA AGT GCG GGA AGA-3'	277
SAP 6	Forward 5'-CCC GTT TTG AAA TTA AAT ATG CTG ATG G-3' Reverse 5'-GTC GTA AGG AGT TCT GGT AGC TTC G-3'	187
SAP 7	Forward 5'-GAA ATG CAA AGA GTA TTA GAG TTA TTA C-3' Reverse 5'-GAA TGA TTT GGT TTA CAT CAT CTT CAA CTG-3'	196

Genes	Infection (n = 11) n (%)	Colonization (n = 15) n (%)	Total (n = 26) n (%)	p-value
SAP1	10 (90.9)	8 (53.33)	18 (69.23)	0.049
SAP2	7 (63.63)	9 (60)	16 (61.54)	0.588
SAP3	5 (45.45)	7 (46.66)	12 (45.15)	0.631
SAP4	10 (90.9)	11 (73.33)	21 (80.77)	0.273
SAP5	8 (72.72)	13 (86.66)	21 (80.77)	0.345
SAP6	8 (72.72)	14 (93.33)	22 (84.61)	0.187
SAP7	11 (100)	11 (73.33)	22 (84.61)	0.091

Pattern	Genotype	n (%)
1	SAP1-7	3 (16)
2	SAP1/SAP2/SAP4/SAP5/SAP6/SAP7	3 (16)
3	SAP1/SAP3/SAP4/SAP5/SAP6/SAP7	2 (10)
4	SAP2/SAP4/SAP5/SAP6	2 (10)
5	SAP2/SAP5/SAP6/SAP7	2 (10)
6	SAP1/SAP2/SAP3/SAP4/SAP5/SAP7	1 (5)
7	SAP1/SAP2/SAP3/SAP5/SAP6/SAP7	1 (5)
8	SAP1/SAP2/SAP3/SAP4/SAP6/SAP7	1 (5)
9	SAP3/SAP4/SAP5/SAP6/SAP7	1 (5)
10	SAP1/SAP3/SAP4/SAP6/SAP7	1 (5)
11	SAP1/SAP2/SAP4/SAP6/SAP7	1 (5)
12	SAP1/SAP2/SAP3/SAP4/SAP7	1 (5)
13	SAP1/SAP4/SAP5/SAP6/SAP7	1 (5)
14	SAP2/SAP3/SAP4/SAP5/SAP6	1 (5)
15	SAP4/SAP5/SAP6/SAP7	1 (5)
16	SAP1/SAP5/SAP6/SAP7	1 (5)
17	SAP1/SAP4/SAP5/SAP7	1 (5)
18	SAP1/SAP4/SAP7	1 (5)
19	SAP5/SAP6	1 (5)

Studies	Population	Frequency of SAP genes
Our study	Women from Rondonópolis with symptomatic VVC	SAP7 (100%) and SAP1-4 (90.9%)
Our study	Women from Rondonópolis asymptomatic for VVC	SAP6 (93.33%) and SAP5(86.66)
Monroy-Pérez et al.⁽³¹⁾	Women from Mexico with VVC	SAP4-6 (100%)
Bassyouni et al.⁽²⁶⁾	Diabetic and non-diabetic women from Egypt	SAP1, SAP2 (100%) and SAP5 (75%)
Kalkanci et al.⁽³²⁾	Women from Turkey with VVC	SAP1-3 (92.5%), SAP6 (12.5%) and SAP4-5 (7.5%)

Study	Population	Expression of SAP genes
Nas et al.⁽⁴¹⁾ Lian and Liu⁽⁴³⁾ Lin et al.⁽⁴⁴⁾	Women from Turkey with VVC Symptomatic, asymptomatic women, and women with RVVC from China Women from China with acute VVC	Expression of SAP4: pregnant (40%), post-menopausal (50%) women, and women in childbearing age (33%) SAP1 and SAP3 were present only in VVC and RVVC isolates SAP2 and SAP5 were most commonly expressed in the vaginal mucosa

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[TFN1] PCR: polymerase chain reaction.