Estimated minimum number of promastigotes of Leishmania (Viannia) braziliensis necessary for proliferation in a culture system in microplates
Keywords:
Microcultivation, L. (V.) braziliensis, Proliferation in vitro, Leishmania, Parasitological diagnosisAbstract
The definitive diagnosis of cutaneous leishmaniasis depends on meeting ofetiological agents. The traditional culture technique (TCT) is developed i n t u b e s d e m a n d p ro l o n g e d i n c u b a t i o n , l a r g e vo l u m e o f t h e cultivationmedium and sample. The microculture technique (MCT) developed in 96 well culture plates use low volume of the cultivation medium and thesample, possible to introduce different inocula per sample examined. Thegrowing of L. (V.) braziliensis (MHOM/84/LTB300) was achieved in the twotechniques to determine the minimum number of the parasites necessaryfor proliferation. From serial dilutions with L. (V.) 0 3 braziliensis, inoculations were obtained at concentrations of 1x10 to 1x10 3 for cultivation. In concentration of 1x10 was observed parasite 0 proliferation since 48 hour of0cultivation. A single parasite (1x10 ) was sufficient to allow the proliferation in both techniques. In the inoculum concentration 1x101 a TTC reached 1x108 parasites, but the proliferation was first observed in the MCT. In concentration of 1x103 the MCT reached 2x108 parasites. It wasconcluded that MCT is a strategy what allowed the cultivation since a singleflagellate form and further studies are being conducted to assess feasibilityof MCT use as a diagnostic method.
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Copyright (c) 2010 Alessandra Gutierres, Luis Felipe Cantini Tolezano, Elizabeth Visone Nunes Westphalen, Roberto Mitsuyoshi Hiramoto, Jeffrey Jon Shaw, José Eduardo Tolezan
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