Abstract. Due to the clinical importance of differentiating the two species of the Entamoeba histolytica/Entamoeba dispar complex, we developed a multiplex polymerase chain reaction (PCR) method that overcomes time-consuming and laborious procedures. We report here a DNA extraction protocol using non-fixed stool samples that avoid long lysis-incubation periods through the combined use of zirconium beads and a lysis-supporting buffer. We characterized 49 of 52 stool specimens from Cuban patients with amoebiosis. Among them, 36 (75.5%) were infected only with E. dispar (the nonpathogenic species), while 13 (24.5%) displayed a mixed infection with both E. dispar and E. histolytica. The multiplex PCR protocol showed a specificity of 1.00 and a sensitivity of 0.94. Furthermore, the entire procedure can be performed in one day. This approach is therefore reliable and applicable in the field for epidemiologic studies.
El sobrediagnóstico microscópico de amebiasis intestinal ha sido ampliamente reportado, pêro poças veces objetivãmente demostrado. En el presente trabajo, empleando Enzymeba, un procedimiento diagnóstico de amebiasis intestinal desarrollado en el Instituto de Medicina Tropical "Pedro Kourí", Ciudad de La Habana, abordamos este problema. Durante seis meses se colectaron cn los servidos de parasitologfa de Ia casi totalidad de los policlínicos (13 de 14), y de los dos mayorcs hospitales (2 de 3) de Ia província de Cienftiegos, todas Ias muestras de heces en Ias que el personal técnico correspondiente había informado Ia detección por observación microscópica de uno o más estádios de Entamoeba histolylica. En solo 61 (14.4%) de 424 muestras colectadas Enzymeba confirmo el hallazgo microscópico prévio. No encontramos diferencia estadísticatnente significativa (p>0.05) entre Ia eficiência dei diagnóstico microscópico en los hospitales y en los policlínicos de aquella província.PALABRAS CLAVES: Amebiasis. Sobrediagnóstico microscópico. Enzymeba.
INTRODUCCIÓNEl examen microscópico de heces continua siendo el procedimiento de laboratório más ampliamente utilizado para el diagnóstico de amebiasis
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