Diagnóstico molecular de cromosomopatías fetales en Costa Rica
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Date
Authors
Malespín Bendaña, Wendy Karina
Ortiz Morales, Fernando
Castro Volio, Isabel
Journal Title
Journal ISSN
Volume Title
Publisher
Acta méd. costarric. Vol 51 (5), octubre-diciembre 2009 p.240-244
Abstract
Justificación y objetivos: En Costa Rica, el diagnóstico de anomalías cromosómicas fetales se
realiza solo mediante el análisis citogenético convencional de cromosomas obtenidos de cultivos
celulares. Además de que la espera por los resultados puede ser larga, con alguna frecuencia
fracasa el cultivo, por contaminación o por mala calidad de la muestra, o las figuras mitóticas no
se pueden analizar, por lo que es necesario disponer de una metodología sencilla y barata, para
obtener un diagnóstico prenatal rápido y fiable de trisomía 21, 18 ó 13, en embarazos de alto
riesgo genético sometidos a amniocentesis o cordocentesis.
Métodos: Se diseñaron tres PCRs multiplex para amplificar cuatro distintas repeticiones cortas
en tándem, de cada uno de los cromosomas 21, 18 y 13. Se colectaron 93 muestras (88 líquidos
amnióticos y 5 sangres fetales), recibidas en el laboratorio entre 2006 y 2008, con solicitud de
análisis cromosómico. Los resultados de la reacción en cadena de la polimerasa cuantitativa
fluorescente, fueron comparados con el cariotipo obtenido de las mismas muestras para demostrar
la fiabilidad del ensayo
Resultados: Para este grupo de datos, la exactitud del ensayo fue del 100% y se consiguió
obtener resultados en 48 horas. Se logró realizar el análisis de repeticiones cortas en tándem en
el 77% de las muestras en las que no se pudo obtener crecimiento celular.
Conclusión: La reacción en cadena de la polimerasa cuantitativa fluorescente demostró ser una
metodología sencilla, fiable y rápida, por lo que podría convertirse en una herramienta
complementaria del análisis cromosómico convencional. La obtención de resultados rápidos en
casos de diagnóstico prenatal podría disminuir el periodo de ansiedad parental por la espera de
los resultados, así como permitir un mejor abordaje terapéutico de los fetos afectados.
Justification and aims: In Costa Rica, the diagnosis of chromosomal fetal anomalies is realized only by conventional cytogenetic analysis of chromosomes obtained from cellular cultures. The waiting for the results can be long. Moreover with some frequency culture fails due to contamination or bad quality of the sample or they cannot be analyzed. This makes it necessary to have a simple and cheap methodology to obtain an accurate and rapid fetal diagnosis of trisomy 21, 18 or 13, in pregnancies of high genetic risk submitted to amniocentesis or cordocentesis.Materials and methods: Three multiplex PCRs were designed to amplify four different short tandem repeats of each of the chromosomes 21, 18 and 13. There were collected 93 samples (88 amniotic fluids and 5 fetal bloods), received in the laboratory between 2006 and 2008 with request ofor chromosomal analysis. The results of the quantitative fluorescent PCR were compared with the obtained cariotype of the same samples to stablish the accuracy demonstrate the reliability of the assay. Results: Accuracy of the assay was 100% and it was possible to obtain results within 48 hours. STRs analysis could be made in 77% of the samples where the cellular culture could not be done. Conclusion: The quantitative fluorescent PCR demonstrated to be a simple, accurate and rapid methodology, from what it might turn into a complementary tool of the chromosomal conventional analysis. The securing of rapid results in cases of antenatal diagnosis might diminish the period of anxiety parental for the waiting of the results, as well as to allow a better therapeutic management of the affected fetuses.
Justification and aims: In Costa Rica, the diagnosis of chromosomal fetal anomalies is realized only by conventional cytogenetic analysis of chromosomes obtained from cellular cultures. The waiting for the results can be long. Moreover with some frequency culture fails due to contamination or bad quality of the sample or they cannot be analyzed. This makes it necessary to have a simple and cheap methodology to obtain an accurate and rapid fetal diagnosis of trisomy 21, 18 or 13, in pregnancies of high genetic risk submitted to amniocentesis or cordocentesis.Materials and methods: Three multiplex PCRs were designed to amplify four different short tandem repeats of each of the chromosomes 21, 18 and 13. There were collected 93 samples (88 amniotic fluids and 5 fetal bloods), received in the laboratory between 2006 and 2008 with request ofor chromosomal analysis. The results of the quantitative fluorescent PCR were compared with the obtained cariotype of the same samples to stablish the accuracy demonstrate the reliability of the assay. Results: Accuracy of the assay was 100% and it was possible to obtain results within 48 hours. STRs analysis could be made in 77% of the samples where the cellular culture could not be done. Conclusion: The quantitative fluorescent PCR demonstrated to be a simple, accurate and rapid methodology, from what it might turn into a complementary tool of the chromosomal conventional analysis. The securing of rapid results in cases of antenatal diagnosis might diminish the period of anxiety parental for the waiting of the results, as well as to allow a better therapeutic management of the affected fetuses.
Description
artículo (arbitrado)--Universidad de Costa Rica. Instituto de Investigaciones en Salud, 2009
Keywords
cromosoma, cromosoma 18, cromosoma 21, Diagnóstico prenatal, Costa Rica, Análisis citogenético, Trisomía, cromosoma 13
Citation
http://actamedica.medicos.cr/index.php/Acta_Medica/article/view/484/450