A rapid PCR-based test for identification of fifteen British bat species

Abstract

The development of a rapid method for identification of the fifteen common species of British bat is described: Barbastella barbastellus, Eptesicus serotinus, Myotis bechsteinii, Myotis daubentonii, Myotis mystacinus, Myotis nattereri, Nyctalus leisleri, Nyctalus noctula, Plecotus auritus, Plecotus austriacus, Pipistrellus nathusii, Pipistrellus pipistrellus, Pipistrellus pygmaeus, Rhinolophus ferrumequinum and Rhinolophus hipposideros. A single multiplex PCR containing five fluorescently-tagged primers and fifteen species-specific primers targeting the mitochondrial control region and genes for cytb and ND1 produces clear peaks that are unique to each species when visualised on a DNA sequencer. The multiplex also includes primers to test for PCR-amplifiable bat DNA. This method will facilitate high throughput species identification of British bats, particularly in cases where phenotypic identification is difficult.