Metarhizium anisopliae is the most popular entomopathogenic fungi used successfully in microbial control. Isolation, identification, and estimation of M. anisopliae Egyptian isolate host range has been done in this study. Our isolate was identified molecularly by amplification of Internal transcribed spacer (ITS) region using polymerase chain reaction technique (PCR). The sequence of ITS was compared with other M. anisopliae isolates published in gene bank and our isolate showed 98% homology with most published M. anisopliae isolates so submitted in gene bank database as a new Egyptian isolate with accession no MT102079.The pathogenicity was evaluated by applying five spore suspension concentrations against four insect pests, Spodoptera littoralis (2^nd and 4^th instar larvae), Galleria mellonella (3^rd and 5^th instar larvae), Tetranychus urticae (nymph) and Rhynchophorus ferrugineus (3^rd and 5^th instar larvae), and our results revealed that M. anisopliae was highly effective against all tested insects. The highest concentration used (10⁹) caused a high mortality rate after 10 days post-treatment reached 96% for T. urticae nymph which appears to be highly sensitive insect tested in this study followed by S, littoralis 2^nd and 4^th instar larvae which showed 96 and 90% mortality rate respectively. High pathogenicity of the tested isolate also noticed in G. mellonella in which the highest concentration used (10⁹) induced 93 and 86% for 3^rd and 5^th instar larvae respectively, while the same instars of R. ferrugineus were moderately affected and 73 and 66% mortality rate was achieved by the same concentration after 10 days post-treatment.