Identification of biomarkers for Miltefosine resistance in Leishmania donovani

To identify biomarkers specific for Miltefosin resistant parasites, we used DNA microarray to identify differentially expressed genes in resistant parasite in comparision to sensitive and further validated with real time PCR assay and immunoblotting assay Clinically proven miltefosine reistant as well as respondant parasite strains isolated from splenic aspirates and grown in M199 media. For microarray resistant and sensitive samples run in duplicate while for real time PCR, 25 resistant and 33 sensitive strains used. For immunoblotting, confirmed genes were cloned and recombinant protein blotted against miltefosine failure and treated patients sera.