EFFECT OF HIGHLY ACTIVE ANTIRETROVIRAL THERAPY ON VAGINAL Candida spp. ISOLATION IN HIV-INFECTED COMPARED TO HIV-UNINFECTED WOMEN

Candidíase vulvovaginal (CVV) em mulheres infectadas pelo HIV contribuiu substancialmente para a diminuição da sua qualidade de vida. O objetivo deste estudo foi avaliar o efeito do uso de terapia anti-retroviral altamente ativa (HAART) no isolamento de Candida spp. vaginais em mulheres HIV positivas comparado às não infectadas por HIV. Este estudo transversal incluiu 178 mulheres infectadas pelo HIV (grupo HIV) e 200 mulheres não infectadas (grupo controle) acompanhadas no Serviço de Assistência Especializada (SAE) para as doenças sexualmente transmissíveis (DST)/AIDS da cidade de Maringá/Brasil, de 1 abril a 30 de outubro de 2011. As leveduras foram isoladas e identificadas por métodos fenotípicos e moleculares. A susceptibilidade in vitro aos antifúngicos fluconazol, itraconazol, nistatina e anfotericina B foi avaliada pelo método de referência de microdiluição. Nós encontramos maior frequência de isolamento vaginal total de Candida spp. no grupo HIV do que no grupo controle. Entretanto, foi observada frequência similar de colonização e CVV entre os dois grupos. Apesar de C. albicans ser a mais frequente e sensível a azólicos e polienos em mulheres infectadas pelo HIV e não infectadas, foi detectada emergente resistência de C. glabrata a AMB nas mulheres infectadas pelo HIV. Embora tenha sido observada maior frequência de isolamento vaginal de Candida spp. nas mulheres infectadas pelo HIV do que nas não infectadas, colonização e CVV apresentaram frequência similar em ambos os grupos, o que indica que HAART parece proteger contra colonização vaginal e CVV. Vulvovaginal candidiasis (VVC) in HIV-infected women contributed to the impairment of their quality of life. The aim of this study was to evaluate the effect of highly active antiretroviral therapy (HAART) use on the vaginal Candida spp. isolation in HIV-infected compared to HIV-uninfected women. This cross-sectional study included 178 HIV-infected (HIV group) and 200 HIV-uninfected women (control) that were studied at the Specialized Assistance Service (SAE) for sexually transmitted diseases (STD)/AIDS of the city of Maringá, Brazil, from April 1 to October 30, 2011. The yeasts were isolated and identified by phenotypic and molecular methods. The in vitro antifungal susceptibility to fluconazole, itraconazole, nystatin and amphotericin B was tested by the reference microdilution method. Higher frequencies of total vaginal Candida spp. isolation were found in the HIV-infected group than in the control group. However, both groups showed a similar frequency of colonization and VVC. Although C. albicans was the most frequent and sensitive to azolics and polyenes in both HIV-infected and uninfected women, the emerging resistance of C. glabrata to amphotericin B in the HIV-infected women was observed. Although higher frequency of vaginal Candida spp. isolation had been observed in the HIV-infected than in HIV-uninfected women, colonization and VVC showed similar frequency in both groups, indicating that HAART appears to protect against vaginal colonization and VVC

Prevalence of Candida albicans and non-albicans isolates from vaginal secretions: comparative evaluation of colonization, vaginal candidiasis and recurrent vaginal candidiasis in diabetic and non-diabetic women

CONTEXT AND OBJECTIVE: Vulvovaginal candidiasis (VVC) is caused by abnormal growth of yeast-like fungi on the female genital tract mucosa. Patients with diabetes mellitus (DM) are more susceptible to fungal infections, including those caused by species of Candida. The present study investigated the frequency of total isolation of vaginal Candida spp., and its different clinical profiles - colonization, VVC and recurrent VVC (RVVC) - in women with DM type 2, compared with non-diabetic women. The cure rate using fluconazole treatment was also evaluated. DESIGN AND SETTING: Cross-sectional study conducted in the public healthcare system of Maringá, Paraná, Brazil. METHODS: The study involved 717 women aged 17-74 years, of whom 48 (6.7%) had DM type 2 (mean age: 53.7 years), regardless of signs and symptoms of VVC. The yeasts were isolated and identified using classical phenotypic methods. RESULTS: In the non-diabetic group (controls), total vaginal yeast isolation occurred in 79 (11.8%) women, and in the diabetic group in 9 (18.8%) (P = 0.000). The diabetic group showed more symptomatic (VVC + RVVC = 66.66%) than colonized (33.33%) women, and showed significantly more colonization, VVC and RVVC than seen among the controls. The mean cure rate using fluconazole was 75.0% in the diabetic group and 86.7% in the control group (P = 0.51). CONCLUSION: We found that DM type 2 in Brazilian women was associated with yeast colonization, VVC and RVVC, and similar isolation rates for C. albicans and non-albicans species. Good cure rates were obtained using fluconazole in both groups

Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR.

Sexually transmitted diseases (STDs) may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR) assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV) -1 and -2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV) and genotypes by single PCR (sPCR) in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%), sensitivity (100.00%), specificity (99.70%), positive (96.40%) and negative predictive values (100.00%) and accuracy (99.80%). The prevalence of STDs was very high (55.3%). Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks

Electrophoretic analysis of the amplified fragments by using a multiplex polymerase chain reaction in 8% polyacrylamide gel stained with ethidium bromide.

<p>Lane C₁: control of <i>Chlamydia trachomatis</i> (361 base pairs-bp); lane C₂: control of <i>Treponema pallidum</i> (291 bp); lane C₃: control of HSV–2 (249 bp); lane C₄: control of <i>Mycoplasma genitalium</i> (193 bp); lane C₅: control of <i>Trichomonas vaginalis</i> (170 bp); lane C₆: control of <i>Neisseria gonorrhoeae</i> (162 bp); lane C₇: control of HSV–1(123 bp); lane A₁: positive sample of <i>C. trachomatis</i> and HSV–1 (361 and 123 bp); lane A₂: positive sample of <i>T. pallidum</i> and HSV–2 (291 and 249 bp); lane A₃: positive sample of <i>T. vaginalis</i> and HSV–2 (170 and 249 bp); lane A₄: positive sample of <i>C. trachomatis</i> and <i>M. genitalium</i> (361 and 193 bp); lane A₅: positive sample of <i>T. pallidum</i> and <i>T. vaginalis</i> (291 and 170 bp); lane A₆: positive sample of <i>T. vaginalis</i> (170 bp); lanes M₁ and M₂, molecular weight marker (25 bp Invitrogen). Values on the left and right sides of the gel are in bp.</p

M–PCR validation results compared with single PCR (sPCR) for seven clinically important STDs pathogens in semen, including <i>Chlamydia trachomatis</i>, <i>Mycoplasma genitalium</i>, <i>Neisseria gonorrhoeae</i>, <i>Treponema pallidum, Trichomonas vaginalis</i>, HSV–2 and HSV–1.

<p>M–PCR, multiplex–polymerase chain reaction; sPCR, single polymerase chain reaction; STDs, sexually transmitted diseases; PPV, positive predictive value; NPV, negative predictive value; ACC, accuracy; CT, <i>C. trachomatis</i>; TP, <i>T. pallidum</i>; HSV–1/–2: herpes virus simplex; MG, <i>M. genitalium</i>; TV, <i>T. vaginalis</i>; NG, <i>N. gonorrhea</i>.</p

Electrophoretic analysis of the HPV genotyping in semen using PCR-RFLP with restriction enzyme <i>Hpy</i>CH4V in 8% polyacrylamide gel stained with ethidium bromide.

<p>Sample A1, genotypes −16 (High-risk, HR) and −31 (HR) in double HPV infection (216, 191, 94 and 91 base pairs-bp); A2, genotype −13 (low-risk, LR) in single HPV infection (244, 103 and 91 bp); A3, genotype −16 (HR) in single HPV infection (216 and 191 bp); A4, genotype −18 (HR) in single HPV infection (174, 144 and 100); A5, genotypes −81(LR), −66 (HR) and −16 (HR) in multiple HPV infection (284, 216, 191 and 89 bp). M, molecular weight marker (25 bp).</p

Frequency of STDs and reason for performing the semen analysis.

<p>STD, sexually transmitted diseases; HPV, human <i>Papillomavirus</i>; HR–HPV, high–risk human <i>Papillomavirus</i>; HSV–1/–2, herpes simplex virus; (+), positive; (−), negative.</p

HPV genotypes distribution of 17 semem samples with two or more genotypes.

<p>HPV, human <i>Papillomavirus</i>; HR, high–risk human <i>Papillomavirus</i>; LR, low-risk human <i>Papillomavirus</i>.</p