The family Spinturnicidae (Oudemans, 1902) comprises hematophagous mites found exclusively on bats. In this article, we present DNA barcodes (mitochondrial Cytochrome c Oxidase I) for 71 specimens morphologically assigned to six spinturnicid species from 34 bat specimens (11 species) obtained from 10 caves and one forest fragment in karst areas from Minas Gerais state, southeast Brazil. A visual guide with diagnostic characters for Periglischrus acutisternus Machado-Allison, 1964, P. caligus Kolenati, 1857, P. herrerai Machado-Allison, 1965, P. iheringi Oudemans, 1902, P. paravargasi Herrin and Tipton, 1975, P. torrealbai Machado-Allison, 1965 is provided. Along DNA barcode data, photo-documentation of vouchering material of each individual sequenced have been deposited at BOLD (Barcoding of Life Data System v4) database. Periglischrus herrerai, P. caligus and P. paravargasi are reported for the first time in Minas Gerais state, extending their distribution ranges.