Please use this identifier to cite or link to this item:
|Title:||High-throughput screening of amastigotes of <i>Leishmania donovani </i>clinical isolates against drugs using a colorimetric β-lactamase assay|
Buckner, Frederick S
Indian clinical isolates
|Abstract:||A simple colorimetric β-lactamase assay for quantifying <i>Leishmania </i>amastigotes in macrophages grown in microtiter plates has been reported. The β-lactamase gene was integrated into the rRNA region of the genome, thereby allowing for high-level stable expression of the enzyme. Both visceral leishmaniasis (VL) and post-kala azar dermal leishmaniasis (PKDL) isolates were transfected with β-Lactamase gene. These β-lactamase-expressing promastigotes were used for infecting intracellular J774A.1 macrophages <i>in vitro</i>. Quantification was done by a colorimetric readout with CENTA<sup>TM</sup> β-lactamase as substrate and with an optical density plate reader. The assay was carried out in 96-well plates. Results obtained demonstrate that this methodology could be a valuable high-throughput screening assay for checking efficacy of anti-leishmanial drugs in the clinical isolates.|
|Appears in Collections:||IJEB Vol.47(06) [June 2009]|
Items in NOPR are protected by copyright, with all rights reserved, unless otherwise indicated.