Volume 49, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



To construct a DNA probe specific for protozoa that cause visceral leishmaniasis, we cloned I fragments of genomic DNA into a Bluescript II SK® vector. A clone of 4.3 kb that contained a highly repetitive sequence was isolated and cut with three restriction enzymes: III, I, and 3A. After a new molecular cloning step, we isolated and sequenced a 140-basepair (bp) fragment. Two oligonucleotides were synthesized to be used as primers for a polymerase chain reaction. Using this probe, we detected an amount of DNA equivalent to one promastigote of . This probe showed a high specificity; all protozoa tested that cause visceral leishmaniasis and (one of the causative agents of Old World cutaneous leishmaniasis) showed a 100-bp amplified sequence, whereas other strains showed a signal of a different size or else no signal. Moreover, no amplified sequence was obtained with other pathogenic parasites tested (, and ).


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