Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina

Introduction. Pregnancy is one of the risk factors for the development of vulvovaginal\ncandidiasis (VVC). Its diagnosis and treatment can prevent potential obstetrical and gynecological\ncomplications. The species Candida albicans is the primary etiological agent; however, VVC due\nto non-Candida a...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autor principal:	Mucci, María Josefina
Otros Autores:	Cuestas, María Lujan
Formato:	Tesis de maestría acceptedVersion
Lenguaje:	Español
Publicado:	Facultad de Farmacia y Bioquímica 2015
Materias:	Candida Albicans Candida no-albicans Embarazadas Candidiasis vulvovaginal CCV Ciencia de la vida
Acceso en línea:	http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1123 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1123.dir/1123.PDF
Aporte de:	Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires

id	I28-R145-HWA_1123
record_format	dspace
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-145
collection	Repositorio Digital de la Universidad de Buenos Aires (UBA)
language	Español
orig_language_str_mv	spa
topic	Candida Albicans Candida no-albicans Embarazadas Candidiasis vulvovaginal CCV Ciencia de la vida
spellingShingle	Candida Albicans Candida no-albicans Embarazadas Candidiasis vulvovaginal CCV Ciencia de la vida Mucci, María Josefina Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina
topic_facet	Candida Albicans Candida no-albicans Embarazadas Candidiasis vulvovaginal CCV Ciencia de la vida
description	Introduction. Pregnancy is one of the risk factors for the development of vulvovaginal\ncandidiasis (VVC). Its diagnosis and treatment can prevent potential obstetrical and gynecological\ncomplications. The species Candida albicans is the primary etiological agent; however, VVC due\nto non-Candida albicans species has increased gradually and is becoming more resistant to\nantifungal agents. Thus, the identification of Candida isolates is essential since conventional\nmethods in diagnostic microbiology are inconclusive for some species.\nObjective. Report on the frequency of VVC and the carriage of Candida species in\nimmunocompetent pregnant patients. Determine the distribution and prevalence of yeasts from\ndifferent species isolated from patients with VVC and asymptomatic carriage of these\nmicroorganisms. Differentiate the C. albicans species complex (C. albicans, C. dubliniensis and C.\nafricana), C. parapsilosis and C. glabrata by molecular methods. Compare conventional\nmicrobiological techniques with molecular methods aimed at its detection. Determine the\nfrequency of other pathogens with increased obstetric risk factors or causing vulovaginal\ninfections and recognize the possible association with the different Candida species.\nMaterials and Methods. We obtained 210 vaginal swabs and 64 vaginal secretion\nsamples from immunocompetent pregnant women at 35-37 weeks of gestation to analyze\nStreptococcus agalactiae and its association with Candida species. For yeast identification,\nconventional microbiological studies were performed (microscopic examination of samples,\nCHROMagar Candida culture media, chlamydoconidia production in milk agar 1% Tween 80 and\nStaib medium, assimilation of sugars, and API ID 32C method) and molecular methods (PCR from\nyeast suspensions using specific primers in order to: i) differentiate between C. albicans, C.\ndubliniensis and C. africana (actin and HWP1 gene); ii) C. parapsilosis, C. orthopsilosis and C.\nmetapsilosis (ITS1-ITS2 regions of the rDNA); and iii) C. glabrata, C. bracarensis and C.\nnivariensis (RLP31 gene encoding a 60S ribosomal protein). Microbiological studies were also\ncarried out by conventional methods for the identification of S. agalactiae, Ureaplasma spp.,\nMycoplasma hominis, Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis.\nStatistical analysis of the results was performed using descriptive statistics and chi-square test;\nGraphPad Prism (version 6.0).\nResults: The prevalence of VVC in pregnant women was 25% (52/210). Increased\nsymptoms were observed as gestation progressed, being higher in the 2nd and 3rd trimester of\npregnancy, probably secondary to the increased colonization by Candida spp. Colonization by\nCandida spp in asymptomatic patients was 13.8%. C. albicans was isolated in 80.7% of VVC\npatients and the remaining 19.3% showed other yeast species, whereas in 50% of asymptomatic\npatients, S. cerevisiae, Rhodotorula spp and non-Candida albicans were isolated (p <0.005). This\nsuggests that yeast colonization in the vagina is associated with non-Candida albicans or other\nyeasts and that the development of VVC could depend, among other factors, on the replacement\nof these species with C. albicans. In the VVC population, the most predominant isolates of non-\nCandida albicans species were C.glabrata (7.68%) and C. dubliniensis (3.84%). The latter two\nspecies were found in pregnant women as well as in colonizing VVC biota, in both cases with no\nsignificant differences.\nThe molecular methods used in this study allowed the differentiation of Candida species not\ndistinguished by conventional phenotyping, like C. parapsilosis sensu stricto. PCR amplification of\nthe intron of the actin gene in C. dubliniensis confirmed the conventional microbiological\nidentification. Neither the presence of C. africana, C. bracarensis nor C. nivariensis could be\nobserved by PCR using specific primers. PCR amplification of the HWP1 gene allowed the\nidentification of isolates of heterozygous and homozygous C. albicans for this gene. No significant\ndifferences were observed in VVC patients or in those with asymptomatic carriage of the\npathogen.\nWhen considering other pathogens engaged in the study population (n=210), we observed that BV\ninvolved 18.09%, C. trachomatis 1.43%, N. gonorrhoeae 0.48%, Ureaplasma spp. and\nMycoplasma hominis 17.62% and 2.85% respectively.\nS. agalactiae was found in 12.5% of pregnant women at 35-37 weeks of gestation (n=64), and\n4.7% was associated with C. albicans.\nSome Candida species were associated with bacterial vaginosis (BV), Streptococcus agalactiae,\nUreaplasma spp. and Mycoplasma hominis.\nConclusions: The prevalence of VVC in pregnant women was 25% with C. albicans being\nthe most common species. The phenotypic methods were inconclusive in identifying the different\nCandida species, while the use of molecular techniques provided a more accurate knowledge of\nthis epidemiology in pregnant women in Argentina, particularly due to the importance of C.\nalbicans and non-C. albicans species.
author2	Cuestas, María Lujan
author_facet	Cuestas, María Lujan Mucci, María Josefina
format	Tesis de maestría Tesis de maestría acceptedVersion
author	Mucci, María Josefina
author_sort	Mucci, María Josefina
title	Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina
title_short	Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina
title_full	Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina
title_fullStr	Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina
title_full_unstemmed	Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina
title_sort	epidemiología molecular de la candidiasis vulvovaginal (cvv) en gestantes en argentina
publisher	Facultad de Farmacia y Bioquímica
publishDate	2015
url	http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1123 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1123.dir/1123.PDF
work_keys_str_mv	AT muccimariajosefina epidemiologiamoleculardelacandidiasisvulvovaginalcvvengestantesenargentina
_version_	1766017451273748480
spelling	I28-R145-HWA_11232019-09-27 Introduction. Pregnancy is one of the risk factors for the development of vulvovaginal\ncandidiasis (VVC). Its diagnosis and treatment can prevent potential obstetrical and gynecological\ncomplications. The species Candida albicans is the primary etiological agent; however, VVC due\nto non-Candida albicans species has increased gradually and is becoming more resistant to\nantifungal agents. Thus, the identification of Candida isolates is essential since conventional\nmethods in diagnostic microbiology are inconclusive for some species.\nObjective. Report on the frequency of VVC and the carriage of Candida species in\nimmunocompetent pregnant patients. Determine the distribution and prevalence of yeasts from\ndifferent species isolated from patients with VVC and asymptomatic carriage of these\nmicroorganisms. Differentiate the C. albicans species complex (C. albicans, C. dubliniensis and C.\nafricana), C. parapsilosis and C. glabrata by molecular methods. Compare conventional\nmicrobiological techniques with molecular methods aimed at its detection. Determine the\nfrequency of other pathogens with increased obstetric risk factors or causing vulovaginal\ninfections and recognize the possible association with the different Candida species.\nMaterials and Methods. We obtained 210 vaginal swabs and 64 vaginal secretion\nsamples from immunocompetent pregnant women at 35-37 weeks of gestation to analyze\nStreptococcus agalactiae and its association with Candida species. For yeast identification,\nconventional microbiological studies were performed (microscopic examination of samples,\nCHROMagar Candida culture media, chlamydoconidia production in milk agar 1% Tween 80 and\nStaib medium, assimilation of sugars, and API ID 32C method) and molecular methods (PCR from\nyeast suspensions using specific primers in order to: i) differentiate between C. albicans, C.\ndubliniensis and C. africana (actin and HWP1 gene); ii) C. parapsilosis, C. orthopsilosis and C.\nmetapsilosis (ITS1-ITS2 regions of the rDNA); and iii) C. glabrata, C. bracarensis and C.\nnivariensis (RLP31 gene encoding a 60S ribosomal protein). Microbiological studies were also\ncarried out by conventional methods for the identification of S. agalactiae, Ureaplasma spp.,\nMycoplasma hominis, Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis.\nStatistical analysis of the results was performed using descriptive statistics and chi-square test;\nGraphPad Prism (version 6.0).\nResults: The prevalence of VVC in pregnant women was 25% (52/210). Increased\nsymptoms were observed as gestation progressed, being higher in the 2nd and 3rd trimester of\npregnancy, probably secondary to the increased colonization by Candida spp. Colonization by\nCandida spp in asymptomatic patients was 13.8%. C. albicans was isolated in 80.7% of VVC\npatients and the remaining 19.3% showed other yeast species, whereas in 50% of asymptomatic\npatients, S. cerevisiae, Rhodotorula spp and non-Candida albicans were isolated (p <0.005). This\nsuggests that yeast colonization in the vagina is associated with non-Candida albicans or other\nyeasts and that the development of VVC could depend, among other factors, on the replacement\nof these species with C. albicans. In the VVC population, the most predominant isolates of non-\nCandida albicans species were C.glabrata (7.68%) and C. dubliniensis (3.84%). The latter two\nspecies were found in pregnant women as well as in colonizing VVC biota, in both cases with no\nsignificant differences.\nThe molecular methods used in this study allowed the differentiation of Candida species not\ndistinguished by conventional phenotyping, like C. parapsilosis sensu stricto. PCR amplification of\nthe intron of the actin gene in C. dubliniensis confirmed the conventional microbiological\nidentification. Neither the presence of C. africana, C. bracarensis nor C. nivariensis could be\nobserved by PCR using specific primers. PCR amplification of the HWP1 gene allowed the\nidentification of isolates of heterozygous and homozygous C. albicans for this gene. No significant\ndifferences were observed in VVC patients or in those with asymptomatic carriage of the\npathogen.\nWhen considering other pathogens engaged in the study population (n=210), we observed that BV\ninvolved 18.09%, C. trachomatis 1.43%, N. gonorrhoeae 0.48%, Ureaplasma spp. and\nMycoplasma hominis 17.62% and 2.85% respectively.\nS. agalactiae was found in 12.5% of pregnant women at 35-37 weeks of gestation (n=64), and\n4.7% was associated with C. albicans.\nSome Candida species were associated with bacterial vaginosis (BV), Streptococcus agalactiae,\nUreaplasma spp. and Mycoplasma hominis.\nConclusions: The prevalence of VVC in pregnant women was 25% with C. albicans being\nthe most common species. The phenotypic methods were inconclusive in identifying the different\nCandida species, while the use of molecular techniques provided a more accurate knowledge of\nthis epidemiology in pregnant women in Argentina, particularly due to the importance of C.\nalbicans and non-C. albicans species. Fil: Mucci, María Josefina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Cuestas, María Lujan Facultad de Farmacia y Bioquímica Mujica, María Teresa Mucci, María Josefina 2015-07-13 Introducción. El embarazo es uno de los factores de riesgo para el desarrollo de una\nCandidiasis vulvovaginal (CVV) y su diagnostico y tratamiento permite prevenir posibles\ncomplicaciones gineco-obstetricas. La especie Candida albicans es la principal agente etiológico,\nsin embargo, las CVV debidas a Candida no- albicans se encuentran en paulatino ascenso y\npresentan una mayor resistencia a los antifúngicos. Por ello, es necesaria la identificación de los\naislamientos de Candida y los métodos convencionales de diagnóstico microbiológico en algunas\nespecies no son concluyentes.\nObjetivo. Informar sobre la frecuencia de las CVV y de la portación por especies de\nCandida en pacientes gestantes inmunocompetentes. Determinar la distribución y prevalencia de\nlas distintas especies de levaduras aisladas en las pacientes con CVV y en la portación\nasintomática de estos microorganismos. Diferenciar mediante técnicas moleculares a las\nespecies de los complejos C. albicans (C. albicans, C. dubliniensis y C. africana), C. parapsilosis\ny C. glabrata. Comparar las técnicas microbiológicas convencionales con las moleculares para su\ndetección. Determinar la frecuencia de otros patógenos de riesgo obstétrico o causantes de\nvulovaginitis y conocer la posible asociación con las diferentes especies de Candida.\nMateriales y Métodos. Se analizaron 210 exudados vaginales obtenidos de gestantes\ninmunocompetentes y 64 secreciones vaginales de embarazadas que se encontraban entre las\nsemanas 35 y 37 de gestación para la investigación de Streptococcus agalactiae y su asociación\ncon especies de Candida. Para la identificación de levaduras se realizaron estudios\nmicrobiológicos convencionales ( estudio microscópico de la muestra, cultivos en medio\nCHROMagar Candida, producción de clamidoconidios en agar leche Tween 80 al 1 % y en medio\nde Staib, asimilación de azúcares, y método API ID 32C) y métodos moleculares (PCR a partir\nde las suspensiones de levaduras con primers específicos para: i) diferenciar entre C. albicans,\nC. dubliniensis y C. africana (gen actina y HWP1); ii) C. parapsilosis, C. orthopsilosis y C.\nmetapsilosis (regiones ITS1-ITS2 del rDNA); y iii) C. glabrata, C. bracarensis y C. nivariensis (gen\nRLP31 que codifica para una proteína ribosomal de 60S). Se realizaron además estudios\nmicrobiológicos por métodos convencionales para la identificación de S. agalactiae, Ureaplasma\nspp., Mycoplasma hominis. Neisseria gonorrhoeae, Chlamydia trachomatis y Trichomonas\nvaginalis, Se realizó el análisis estadístico de los resultados mediante estadísticas descriptivas y\nel test de Chi cuadrado. Programa GraphPad Prism (versión 6.0).\nResultados: La prevalencia de CVV en las gestantes estudiadas fue del 25% (52/210). Se\nobservó un incremento de los síntomas a medida que aumentaron las semanas de gestación,\nsiendo mayor en el 2do y 3er trimestre del embarazo, probablemente secundario a la mayor\ncolonización por Candida spp. La colonización por Candida spp alcanzó en las pacientes\nasintomáticas el 13,8 %. En las pacientes con CVV se aisló C. albicans en el 80,7% y el 19,3%\nrestante fueron otras especies de levaduras, en tanto que en las pacientes asintomáticas en el\n50% se aislaron S. cerevisiae, Rhodotorula spp y Candida no- albicans (p <0,005). Ello sugiere\nque la colonización de levaduras en la vagina se asocia con Candida no- albicans u otras\nlevaduras y que el desarrollo de la CVV dependería entre otros factores del reemplazo de estas\nespecies por C. albicans. En las CVV, los aislamientos de especies de Candida no- albicans\ndominantes fueron C. glabrata (7,68%) y C. dubliniensis (3,84%). Estas dos últimas especies, se\nhallaron en las gestantes tanto como biota colonizante como en las VVC, en ambos casos con\nuna diferencia no significativa.\nLos métodos moleculares empleados en el presente trabajo permitieron diferenciar las especies\nde Candida no distinguidas mediante métodos fenotípicos convencionales, como a C.\nparapsilosis sensu stricto. La amplificación por PCR para el intrón del gen de actina en C.\ndubliniensis corroboró la identificación microbiológica convencional. No se registró mediante la\nPCR con los cebadores específicos la presencia de C. africana, C. bracarensis y C. nivariensis.\nLa amplificación por PCR del gen HWP1 permitió detectar aislamientos de C albicans\nheterocigotos y homocigotas para este gen, no encontrándose diferencias significativas en las\npacientes con CVV y en aquellas con portación asintomática de este patógeno.\nAl considerar otros patógenos en la población estudiada (n=210) observamos que la VB afectó\nal 18,09 %, C. trachomatis al 1,43%, N. gonorrhoeae al 0,48%, Ureaplasma spp. y Mycoplasmas\nhominis al 17,62% y 2,85%, respectivamente.\nEn las pacientes que cursaban el embarazo entre las semanas 35 y 37 (n= 64) se halló S.\nagalactiae en el 12,5 % de las pacientes, y en un 4,7% se asoció con C. albicans.\nAlgunas especies de Candida se asociaron con vaginosis bacteriana (VB), Streptococcus\nagalactiae, Ureaplasma spp. y Mycoplasma hominis.\nConclusiones: La prevalencia de CVV en embarazadas fue del 25% y C. albicans la\nespecie más frecuente. Los métodos fenotípicos no resultaron concluyentes en la identificación\nde las distintas especies de Candida, mientras que el empleo de técnicas moleculares, permitió\nun conocimiento más certero de la epidemiología en mujeres gestantes en Argentina,\nparticularmente por la importancia en especies C. albicans y no albicans. application/pdf Catalano, Mariana Arechavala, Alicia Vay, Carlos Candida Albicans Candida no-albicans Embarazadas Candidiasis vulvovaginal CCV spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencia de la vida Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina info:eu-repo/semantics/masterThesis info:ar-repo/semantics/tesis de maestría info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1123 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1123.dir/1123.PDF

Epidemiología molecular de la candidiasis vulvovaginal (CVV) en gestantes en Argentina

Ejemplares similares