Rapid identification of non-neosomic Tunga penetrans and Tunga trimamillata (Insecta Siphonaptera) specimens through PCR-RFLP method

Tunga penetrans (L.) and T. trimamillata Pampiglione et al. are the unique species of the genus parasiting men. While morphological identification of gravid females is possible, the diagnosis on non-gravid females, males or body fragment is quite difficult, time consuming and often requires taxonomic experience. In this paper we present a protocol for the diagnosis of these species based on the PCR-RFLP method. We used the ribosomal intergenic spacer 2 (ITS2) region that was invariable within species but variable among the two taxa. PCR-RFLP confirmed DNA sequence data and demonstrate the reliability of this rapid and relatively inexpensive method to unambiguously discriminate T. penetrans and T. trimamillata.