Abstract: Direct sampling, detection and quantification of entomopathogens can directly reflect the epizootiology of pest in abundance. Pandora neoaphidis, a widespread obligated-aphid pathogen, was investigated in aphid populations. A PCR technology system based on modified Lysis extraction associated with Chelex-100 was developed for epizootiological investigation of entomophthoralean fungi on wheat aphid. Results show that the developed method not only provide a rapid and accurate tool for epizootiological investigation of entomopathogenic fungus, but also have a potential application for rapid and tiny-nucleic acid detection in other fungus.