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

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

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

ABSTRACT

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.⁽³¹31 Monroy-Pérez E, Paniagua-Contreras G, Vaca-Paniagua F, Negrete- Abascal E, Vaca S. SAP expression in Candida albicans strains isolated from Mexican patients with vaginal candidosis. Int J Clin Med. 2013; 4: 25-3.⁾	Women from Mexico with VVC	SAP4-6 (100%)
Bassyouni et al.⁽²⁶26 Bassyouni RH, Wegdan AA, Abdelmoneim A, et al. Phospholipase and aspartyl proteinase activities of Candida species causing vulvovaginal candidiasis in patients with type 2 diabetes mellitus. J Microbiol Biotechnol. 2015; 25(10): 1734-41.⁾	Diabetic and non-diabetic women from Egypt	SAP1, SAP2 (100%) and SAP5 (75%)
Kalkanci et al.⁽³²32 Kalkanci A, Bozdayi G, Birib A, Kustimur S. Distribution of secreted aspartyl proteinases using a polymerase chain reaction assay with SAP specific primers in Candida albicans isolates. Folia Microbiol. 2005; 50(5): 409-13.⁾	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.⁽⁴¹41 Nas T, Kalkanci A, Ozkan S. Expression of ALS1 and SAP4 genes in Candida albicans strains isolated from women with vaginitis. Folia Microbiol. 2008; 53(2): 179-83.⁾ Lian and Liu⁽⁴³43 Lian CH, Liu WD. Differential expression of Candida albicans secreted aspartyl proteinase in human vulvovaginal candidiasis. Mycoses. 2007; 50: 383-90.⁾ Lin et al.⁽⁴⁴44 Lin N, Feng J, Tu Y, Feng A. Expression of Candida albicans secreted aspartyl proteinase in acute vaginal candidiasis. J Huazhong Univ Sci Technol Med Sci. 2007; 27(3): 333-5.⁾	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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[1] CORRESPONDING AUTHOR. Letícia Silveira Goulart, Avenida dos Estudantes, 5005; Cidade Universitária; CEP: 78735-901; Rondonópolis-MT, Brasil; e-mail: lgoulart77@yahoo.com.br.