Alternative PCR protocol using a single primer set for assessing DNA quality in several tissues from a large variety of mammalian species living in areas endemic for leishmaniasis
Mem. Inst. Oswaldo Cruz
;
105(7): 895-898, Nov. 2010. ilus, tab
Artigo
em Inglês
| LILACS
| ID: lil-566179
ABSTRACT
The aim of this work was to establish a modified pre-diagnostic polymerase chain reaction (PCR) protocol using a single primer set that enables successful amplification of a highly conserved mammalian sequence in order to determine overall sample DNA quality for multiple mammalian species that inhabit areas endemic for leishmaniasis. The gene encoding interphotoreceptor retinoid-binding protein (IRBP), but not other conserved genes, was efficiently amplified in DNA samples from tail skin, ear skin, bone marrow, liver and spleen from all of the species tested. In tissue samples that were PCR-positive for Leishmania, we found that DNA from 100 percent, 55 percent and 22 percent of the samples tested resulted in a positive PCR reaction for the IRBP, beta-actin and beta-globin genes, respectively. Nucleotide sequencing of an IRBP amplicon resolved any questions regarding the taxonomical classification of a rodent, which was previously based simply on the morphological features of the animal. Therefore, PCR amplification and analysis of the IRBP amplicon are suitable for pre-diagnostically assessing DNA quality and identifying mammalian species living in areas endemic to leishmaniasis and other diseases.
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Proteínas de Ligação ao Retinol
/
Leishmaniose
/
Reação em Cadeia da Polimerase
/
DNA de Protozoário
/
Actinas
/
Proteínas do Olho
/
Globinas beta
Tipo de estudo:
Guia de Prática Clínica
Limite:
Animais
Idioma:
Inglês
Revista:
Mem. Inst. Oswaldo Cruz
Assunto da revista:
Medicina Tropical
/
Parasitologia
Ano de publicação:
2010
Tipo de documento:
Artigo
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