Molecular methods have revolutionized insect systematic in Metarhizium genera. The importâãncia these entomopathogenic fungus in agriculture is enormous. . In Brazil, to control the sugar cane spittlebug, Mahanarva posticata (Homoptera: Cercopidae). Another very important factor for the selection of isolates of M. anisopliae for use in biological control of pests are particular virulence, reproductive aspects, the media of artificial culture and that the genetic variability of these entomopathogenic fungi have great potential for pest control and the lack of a link between isolate-host-site with the ability of fungal virulence. We studied 5 Metarhizium host (sugar cane spittlebug (IPA213, IPA215, IPA219 and IPA216) and the grasshopper (IPA217). The results obtained with the ITS1 and ITS4 markers showed that these isolates although they infect the same host, are genetically different, perhaps because of their origin as they do not know the origin of insects. However, the isolate IPA217 despite being isolated from grasshopper did not behave differently from the others showing that M. anisopliae complex is monophyletic. We conclude that the greater use of markers and morphological studies can yield good results that may corroborate the idea that this genus investigated phenotypic and molecular characteristics for its identification and typing a new species of Metarhizium.