La rapidité du diagnostic de candidémie et de candidose invasive (CI) est cruciale pour permettre l'introduction précoce des antifongiques. Cette dernière est souvent retardée car l’ hémoculture, test diagnostique de référence, n’est positive que dans 50% des cas avec un délai de plusieurs jours. Des méthodes complémentaires consistent en la détection sérique de polysaccharides de la paroi des Candida : les β-glucanes (BDG) et les mannanes (Man). Nos objectifs étaient de : a) évaluer la signification et l’intérêt clinique de la détection des BDG et des Man en réanimation b) de participer à l’analyse de l’intérêt d’une méthode de spectrometrie de masse (MS) en développement visant à détecter /caractériser des glycannes fongiques circulants. Matériels et Méthodes : a) Pour l'étude clinique cas-contrôles, la cinétique des BDG/Man par rapport à la candidémie a été évaluée chez 41 patients candidémiques et a été comparée à celle relevée chez 67 patients non candidémiques, hospitalisés dans le même service et chez qui la colonisation était déterminée toutes les semaines. b) une méthode de type MALDI-MS a été appliquée à des sérums après extraction sélective des glycannes et comparée à des standards. Resultats : au seuil recommandé le BDG est un biomarqueur sensible, précoce, mais peu spécifique pour le diagnostic de candidémie en réanimation. Il ne semble pas affecté par la colonisation mais sa décroissance lente le rend peu utile pour le suivi du traitement. A l'opposé, le Man est un biomarqueur très spécifique mais peu sensible. En changeant les seuils du BDG, son utilisation séquentielle avec le Man permet de proposer un algorithme dans le cadre d'une stratégie thérapeutique préemptive. b) La méthode MALDI-MS permet de révéler un signal m/z 365 correspondant à un disaccharide (dont le tréhalose) associé aux CI humaines et expérimentales. Des études préliminaires montrent que ce test semble avoir une bonne sensibilité et une bonne spécificité pour le diagnostic de CI ainsi que pour d’autres infections fongiques. Conclusion : a) Le BDG est un marqueur sensible, le Man est une marqueur spécifique, leur utilisation conjointe peut permettre un diagnostic précoce des CI et une rationnalisation du traitement antifongique. b) L'application de la méthode MALDI-MS à la détection/caractérisation des glycannes circulant semble présenter un important potentiel pour se substituer ou complémenter les méthodes diagnostiques actuelles de CI. / The rapidity of the diagnosis of candidaemia and invasive candidiasis (IC) is crucial to allow the early introduction of antigungal therapy. This one is often delayed because Candida yeasts are found in blood culture, the gold standard diagnostic test, in only 50% of cases of IC and several days are needed to have this result. Complementary methods relie on the detection of Candida cell wall polysaccharids in the serum, β-glucans (BDG) and mannans (Man). Our objectives were to : a) determine the signification and clinical interest of the detection of BDG and Man in intensive care b) take part in the analysis of the interest of a mass spectrometry (MS) technic in development which aim to detect/caracterize circulant fongic glycans. Materials and Methods : a) For the clinical case-control study, the BDG and Man kinetics in relation with candidaemia were evaluated in 41 candidemic patients, and were compared to the kinetic observed in 67 non candidemic patients, hospitalised in the same ward and assessed weekly for yeast colonisation b) a MALDI-MS method was applied to sera after selective extraction of glycans and compared to standards. Results : BDG at the recommanded cut-off is a sensitive and precoce but non specific test for the diagnosis of candidemia in ICU. It does not seem to be affected by the colonisation, but its very slow decrease limits its usefulness for the management of the treatment. At the opposite, Man is very specific but not sensitive. Modulating the cut-off of BDG, it is possible to propose a decisional algorithm for preemptive therapy based on the sequential use of BDG and Man. b) MALDI-MS reveals a signal at position m/z 365, corresponding to a disaccharid (among which is trehalose), associated to human and experimental IC. In preliminary studies this test seems to have good sensitiity and specificity for the diagnosis of IC as well as for other fungal infections. Conclusion : a) BDG is a sensitive test, Man is a specific one, and their joint use could be useful for an early diagnosis of IC and a rationalization of the antifungal treatment. b) The application of MALDI-MS method to the detection/caracterisation of circulating glycans seems to present an important potential to replace or complete the current available diagnostic tools of IC.

Orientadores: Paulo Cesar Giraldo, Ana Katherine S. Gonçalves / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-08T15:31:40Z (GMT). No. of bitstreams: 1 Amaral_RoseLuceGomesdo_M.pdf: 734908 bytes, checksum: 93d7f5663377e139f755920fc4edece2 (MD5) Previous issue date: 2007 / Resumo: Objetivo: Verificar se o uso de duchas higiênicas íntimas (DHI) pode interferir na microbiota vaginal de mulheres profissionais do sexo. Sujeitos e Métodos: Estudo de corte transversal realizado durante seis meses verificou o risco de 155 mulheres profissionais do sexo (PS), usuárias e não usuárias de DHI, apresentarem flora vaginal alterada e/ou vaginose bacteriana. As PS foram atendidas em centro de saúde (CS) localizado em zona de prostituição no município de Campinas, São Paulo, e agrupadas em usuárias e não usuárias de duchas higiênicas íntimas para análise da microbiota vaginal. A anamnese determinou os perfis sociodemográfico e sexual destas mulheres que procuraram o CS por diferentes motivos. O conteúdo vaginal foi coletado com swab estéril de Dacron e disposto em duas lâminas de vidro que foram coradas por técnica de Gram. A caracterização da microbiota vaginal pelos critérios de AMSEL adaptado foi realizada por microscopia óptica com lente de imersão. Análises univariável, bivariável com testes de qui-quadrado e exato de Fisher, além de regressão múltipla variável (stepwise) demonstraram o poder de associação entre as variáveis. Resultados: A média de idade, a etnia branca, a escolaridade básica e hábito de fumar encontrados respectivamente nas 94 usuárias e nas 61 não usuárias de DHI foram de 25,5 (± 6,2) vs. 26,0 (± 6,8)anos (p=ns), 48,9% vs. 47,5% (p=ns), 60,6% vs. 45% (p=ns) e 41,5% vs. 49,2% (p=ns) dos casos. Não houve diferença no uso regular de condom com seus parceiros sexuais fixos. Apenas o uso de lubrificantes vaginais foi significativamente maior nas usuárias de DHI (63,8%) que nas não usuárias (36,1%), p=0,0007. As prevalências de flora vaginal alterada, vaginose bacteriana, candidíase, tricomoníase e vaginose citolítica foram de 75,48%; 50,96%; 5,1%; 0,64% e 1,9%, respectivamente. Não houve diferenças significativas destas prevalências quando foram analisados estes achados entre as usuárias e não usuárias de DHI (78,7% vs. 70,5%, ns), (47,9% vs. 55,7%, ns), (5,3% vs. 4,9%, ns), (0 vs. 1,6%, ns) e (1,1% vs. 3,3%, ns). A análise de regressão múltipla tipo stepwise não identificou qualquer risco aumentado de ter flora vaginal alterada ou de vaginose bacteriana em mulheres usuárias de DHI. Conclusão: O uso de DHI não aumentou o risco de flora vaginal alterada e/ou de vaginose bacteriana em mulheres PS. Palavras-chave: profissionais do sexo, duchas higiênicas íntimas, microbiota vaginal, vaginose bacteriana, candidíase vaginal / Abstract: Aims: Verify if vaginal douching (VD) can cause vaginal flora imbalance in Female Sex Workers (FSW). Patients and Methods: A cross sectional study, carried out for six months, analysed the risk of vaginal flora imbalance and/or vaginal bacteriosis due to vaginal douching in 155 FSW. The FSW were seen at a public outpatient in a prostitution area in the city of Campinas, the state of São Paulo, Brazil. The vaginal douching (VD) users and non-users made up the two groups for vaginal microbiological analysis. The social-demographic and sexual profile of these women, who were seen for different reasons, was checked by anamnesis. Vaginal content was collected using a sterile Dacron Swab and placed on two glass slides that were stained using Gram technique. Adapted AMSEL criteria characterized the vaginal flora using optic microscope with immersion lens. Univariate, bivariate statistical analyses with X2 and exact Fisher tests apart from multiple variate regression (Stepwise) analysis determined the association between variables. Results: Mean age, white ethnicity, level of schooling and the habit of smoking analysed in the 94 users and 61 non-users of VD respectively were 25.5 (± 6.2) vs. 26.0 (± 6.8) years (p=ns), 48.9% vs. 47.5% (p=ns), 60.6% vs. 45% (p=ns) and 41.5% vs. 49.2% (p=ns) of the cases. There was no difference in the regular use of condoms with their steady sexual partners. Only the use of vaginal lubricant was significantly higher in the VD users (63.8%) than in the non-users (36.1%); p=0.0007. General prevalence of abnormal flora, bacterial vaginosis, candidiasis, trichomoniasis and cytolitic vaginosis was 75.48%, 50.96%, 5.1%, 0.64% and 1.9% respectively. There were no significant differences when analysing the findings between users and non-users of VD (78.7% vs. 70.5%; ns); (47.9% vs. 55.7%; ns); (5.3% vs. 4.9%; ns); (0 vs. 1.6% ns) and (1.1% vs. 3.3%; ns). Stepwise multiple regression analysis did not identify any increased risk of altered vaginal flora or bacterial vaginosis in VD users. Conclusion: The use of VD did not increase the risk of vaginal flora imbalance and/or bacterial vaginosis in FSW / Mestrado / Ciencias Biomedicas / Mestre em Tocoginecologia