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An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure
Rodríguez, Islay; Fraga, Jorge; Noda, Angel Alberto; Mayet, Milagro; Duarte, Yanisia; Echevarria, Eduardo; Fernández, Carmen.
  • Rodríguez, Islay; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
  • Fraga, Jorge; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
  • Noda, Angel Alberto; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
  • Mayet, Milagro; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
  • Duarte, Yanisia; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
  • Echevarria, Eduardo; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
  • Fernández, Carmen; Instituto de Medicina Tropical Pedro Kourí. La Habana. CU
Braz. arch. biol. technol ; 57(4): 542-547, Jul-Aug/2014. tab, graf
Artículo en Inglés | LILACS | ID: lil-712930
ABSTRACT
Four variants of the potassium acetate procedure for DNA extraction from ixodid ticks at different stage of their life cycles were evaluated and compared with phenol-chloroform and ammonium hydroxide methods. The most rapid and most efficient variant was validated in the DNA extraction procedure from the engorged ticks collected from bovine, canine as well as from house ticks for the screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. The ammonium hydroxide procedure was used for non-engorged ticks. All the variants were efficient and allowed obtaining PCR-quality material according to the specific amplification of 16S rRNA gene fragment of the original tick. DNA extracted from the ticks under the study was tested by multiplex PCR for the screening of tick-borne pathogens. Anaplasma spp. and Babesia spp. amplification products were obtained from 29/48 extracts. Ammonium hydroxide protocol was not efficient for two extracts. Detection of amplification products from the PCR indicated that DNA had been successfully extracted. The potassium acetate procedure could be an alternative, rapid, and reliable method for DNA extraction from the ixodid ticks, mainly for poorly-resourced laboratories.


Texto completo: Disponible Índice: LILACS (Américas) Idioma: Inglés Revista: Braz. arch. biol. technol Asunto de la revista: Biologia Año: 2014 Tipo del documento: Artículo País de afiliación: Cuba Institución/País de afiliación: Instituto de Medicina Tropical Pedro Kourí/CU

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Texto completo: Disponible Índice: LILACS (Américas) Idioma: Inglés Revista: Braz. arch. biol. technol Asunto de la revista: Biologia Año: 2014 Tipo del documento: Artículo País de afiliación: Cuba Institución/País de afiliación: Instituto de Medicina Tropical Pedro Kourí/CU