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Título:	Discrimination of viable acanthamoeba castellani trophozoites and cysts by propidium monoazide real-time polymerase chain reaction
Autor/a:	Fittipaldi Gustavino, Mariana; Pino Rodriguez, Nancy J.; Adrados Ruiz, Barbara; Agustí Adalid, Gemma; Peñuela, Gustavo; Morató Farreras, Jordi; Codony Iglesias, Francesc
Otros autores:	Universitat Politècnica de Catalunya. Departament d'Òptica i Optometria; Universitat Politècnica de Catalunya. TMAS (T+) - Toxicologia i Microbiologia Ambiental i Sanitària
Abstract:	Even though the advent of quantitative polymerase chain reaction (PCR) has improved the detection of pathogen microorganisms in most of areas of microbiology, a serious limitation of this method may arise from the inability to discriminate between viable and nonviable pathogens. To overcome it, the use of real-time PCR and selective nucleic acid intercalating dyes like propidium monoazide (PMA) have been effectively evaluated for different microorganisms. To assess whether PMA pretreatment can inhibit PCR amplification of nonviable amoeba DNA, Acanthamoeba castellani survival was measured using cell culture and real-time PCR with and without PMA pretreatment. Autoclave and contact lens disinfecting solutions were used to inactivate amoebae. After these inactivation treatments, the results indicated that the PMA pretreatment approach is appropriate for differentiating viable A. castellani, both trophozoites and cysts. Therefore, the PMA-PCR approach could be useful as a rapid and sensitive analytical tool for monitoring treatment and disease control, assessing effective disinfection treatments, and for a more reliable understanding of the factors that contribute to the interaction amoeba– pathogenic bacteria.
Abstract:	Peer Reviewed
Materia(s):	-Àrees temàtiques de la UPC::Ciències de la visió::Biologia ocular -Contact lens solutions -Lents de contacte -- Biomicroscòpia
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Tipo de documento:	Artículo - Versión publicada Artículo
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