Multicentric gliomas are rare lesions of central nervous system. Due to the invasive nature of glioma, differentiating multicentric gliomas from diffuse gliomas is difficult. As yet there remains lack of methods to elucidate lesions are totally separated unless surgical biopsy. We use probabilistic fiber tracking to identify the white matter dissemination routes. Our results showed the connectivity value and probabilistic value of the lesions in multicentric gliomas were significantly lower than the lesions in diffuse gliomas. The features provide new insight into multicentric gliomas and may aid in accurate classification of multicentric glioma and diffuse glioma in vivo.