Brugia malayi is a filarial nematode causing lymphatic filariasis in humans characterized by swelling of the lower extremities. The aim of this study was to conduct a real time PCR (qRT-PCR) to verify gene expression levels of Brugia malayi nematodes treated with 20 hydroxyecdysone. Transcriptome analysis was previously performed resulting in the identification of 44 genes that were upregulated by exposure to 20-hydroxyecdysone. Based on transcriptome results, known GO Terms and functions, four genes and one endogenous housekeeping gene were chosen for validation by RT-PCR. Induced samples showed a mean increase of microfilarie by 2.2 fold. Induced wells exhibited a 2.8 fold increase of pre- microfilarie production. On day two adult females treated with 20-HE displayed 3.8-fold increase of microfilaria production as compared to uninduced controls. Overall, all four genes showed upregulation with treatment of 20-hydroxyecdysone at levels that corresponded to the results obtained from the transcriptome analysis. Findings in this experiment expand on the understanding of the ecdysone response system in Brugia malayi, which could serve as a potential drug target against filarial disease.