Variabilidad molecular de aislamientos de Candida spp.por la técnica de polimorfismos de ADN amplificados aleatoriamente (Random Amplified Polymorphic DNA, RAPD) en mujeres de Armenia, Colombia

• Autores: Alba Cecilia Ruiz, Diana Milena Calderón, Jorge E. Gómez Marín
• Localización: Infectio: Revista de la Asociación Colombiana de Infectología, ISSN-e 2422-3794, ISSN 0123-9392, Vol. 13, Nº. 1, 2009
• Idioma: español
• Títulos paralelos:
  • Molecular variation of Candida spp. isolates using Random Amplified Polymorphic DNA (RAPD), from women in Armenia, Colombia
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• Resumen
  • español

    Objetivos. Caracterizar las especies de Candidaque colonizan el conducto vaginal y surelación con sintomatología en las pacientesa quienes se les hace citología vaginal en Armenia(Quindío) y establecer el mejor cebadorpara la técnica de amplificación aleatoria deADN polimórfico (Random Amplified PolymorphicDNA, RAPD).Materiales y métodos. Se recolectaron 226muestras de flujo vaginal de mujeres que asistíana citología rutinaria preventiva y se les realizócultivo en medio de Sabouraud, prueba detubo germinal y batería de asimilación de carbohidratos.Se probaron tres cebadores para laamplificación aleatoria de ADN polimórfico.

  • English

    Objectives: To characterize the Candida species in samples of routine vaginal cytology in Armenia (Colombia) and their relationship with symptoms. Also, to select the adequate primer for a random amplified polymorphismc DNA analysis (RAPD) applied to the Candida species to determine the level of genetic relation.Materials and methods: A total of 226 samples of vaginal secretions were obtained in patients that assist for routine preventive vaginal cytology. The samples were cultured in Sabouraud and then germ tube assay and carbohydrates assimilation test were performed. Three primers were assayed to determine the best for a reproducible RAPD technique.Results: The prevalence of Sabouraud positive cultures was 31.8% (77 of 226 samples) and Candida albicans was the predominant species, representing 71% (55 de 77 samples). Vaginitis symptoms were more frequent in women with positive Sabouraud cultures (93%) vs. women with negative Sabouraud cultures (37%). The best extraction method for DNA extraction was freezing and thawing cycles followed by enzymatic lyses with lyticase. In 47 isolates of Candida species, the best primer that resulted in significant polymorphisms was the OPA 9. A 95% similarity coefficient was obtained 6 clusters. Clusters IV and V only included C. albicans isolates producing symptoms. In cluster V all the isolates were from the same consultation center.Conclusions: C. albicans was the predominant yeast in Armenia and was related with a greater frequency of symptoms. The RAPD technique with OPA9 primer in Armenia, unravel genetic relations of C. albicans isolates that could be used for epidemiological studies.