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Am J Trop Med Hyg ; 49(3): 348-56, 1993 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-8396860

RESUMEN

Oligonucleotide primers directed to the minicircle kinetoplast DNA of Leishmania strains supported enzymatic amplification of this DNA by the polymerase chain reaction (PCR). A single product of 70 basepairs was obtained from parasites belonging exclusively to the L. braziliensis complex. Direct sequencing of the amplified product confirmed its minicircle origin. Skin biopsy specimens from human patients were used directly for the PCR. A pulse incubation of such specimens with deoxyribonuclease I prior to the PCR increased the reliability of the assay. Nuclease disruption of the kinetoplast network was expected to make more copies of the minicircle accessible to amplification. Comparative results between the PCR and conventional parasitologic detection procedures indicate that the DNA detection approach presented is by far more sensitive for diagnostic purposes. Innovations in the PCR protocol are presented that adapt the diagnosis of leishmaniasis to settings with minimal equipment and that are distant from central laboratories.


Asunto(s)
ADN Circular/análisis , Leishmania braziliensis/genética , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Mucocutánea/diagnóstico , Reacción en Cadena de la Polimerasa , Animales , Secuencia de Bases , ADN de Cinetoplasto , ADN Protozoario/análisis , Estudios de Evaluación como Asunto , Humanos , Datos de Secuencia Molecular , Oligodesoxirribonucleótidos/química , Sensibilidad y Especificidad , Piel/parasitología
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