The aim of the present study was to investigate the antibacterial properties of Acalypha indica. Number of resistant antibiotics is increasing and antibacterial activity testing is one of the steps to find treatment of diseases. Acalypha indica is well known medicinal plant and it is a wild plant. It was used as infection treatment for generation to treat several diseases such as asthma, and pneumonia.The susceptibility of five human pathogenic bacterial species to Acalyphin isolated from the leaves of Acalypha indica plant was screened using the agar well diffusion and broth micro-dilution assay. The purity of Acalyphin isolated was checked by TLC and column chromatography and total Acalyphin were quantified. In the present study, the inhibitory action of the Acalyphin was found to increase with an increase in concentration against all bacterial strains. The maximum zone of inhibition was observed at the concentration of 500 µg/ml against all the bacteria. In this study, the S. aureus and E. coli are the more susceptible than the other selected human pathogenic bacteria. From the above investigation the experimental Acalypha indica plant may solve the multidrug resistant bacteria problem and further higher studies is need for qualitative study for the present investigation.