The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) is the major mite pest infesting many cultivated crops. The fungus, Aspergillus aflatoxiformans Frisvad, Ezekiel, Samson & Houbraken produces aflatoxins where it considered as secondary polyketide metabolites, which cause the death of pests. Therefore, this work aimed to isolate and identify this fungus by amplify the internal transcribed spacer (ITS) region of the rDNA, as well as to evaluate the efficiency of this fungus to control T. urticae.  Results showed 300bp DNA product corresponding to the omt-A gene. The existence of omt-A gene in A. aflatoxiformans reflected its ability to produce aflatoxin, which may be responsible of mite control.  The efficacy of A. aflatoxiformans against T. urticae recorded the highest reduction percentage of eggs hatch at concentration of 1x10⁸ conidia/ml within the 5^th and 6^th day after exposure. These percentages were 99.42 and 100%, respectively. The highest mortality percentage was recorded on the 4^th, 5^th and 6^th day at higher concentration of 1x10⁸. No eggs and larvae of T. urticae were observed within the 5^th and 6^th day after treatment at 1x10⁸. There were not observed nymphs at all tested concentrations. Microelectronic photos showed fungal conidia with cluster shaped which were developed at all T. urticae body surface 6 days after treatments at 1x10⁷ and 1x10⁸. Results also confirmed that A. aflatoxiformans could be used successfully to control T. urticae eggs, immature stages and adults. Therefore, this fungus could be involved in any IPM programs of the two-spotted mite.