Abstract: Leaf samples of Lawsonia inermis were collected from Dammar reigon, north of Sudan to examine their antimicrobial potential. Water, methanol and chloroform crude extracts in different concentrations were obtained and bioassayed in vitro for its bioactivity to inhibit the growth of 6 human pathogenic fungi and 4 types of bacteria. The differences in bioactivity of the 3 types of extracts were analyzed. Despite extreme fluctuations in activity, the extract of water was clearly superior. Then followed by methanol, while chloroform extract showed the least. The growth of all pathogens was inhibited to varying degrees by increasing the concentration of the extract. Phytochemical analyses showed the presence of anthraquinones as major constituents of the plant leaves and are commonly known to posses antimicrobial activity. These results confirm the antibacterial and antifungal activity of henna leaves and support the traditional use of the plant in therapy of bacterial infections. The possibility of therapeutic use of Sudanese henna as antimicrobial agents is worthy of note.