As part of a respiratory pathogen survey of Alaska wildlife, we conducted a concordance study to assess Mycoplasma ovipneumoniae detection among three different PCR assays using a total of 346 nasal swabs sampled from four species (Dall's sheep, Ovis dalli dalli; mountain goats, Oreamnos americanus; caribou, Rangifer tarandus granti; and moose, Alces alces gigas), and two taxonomic subfamilies (Bovidae subfamily Caprinae and Cervidae subfamily Capreolinae). A federal research laboratory performed two PCR assays (LM40 and intergenic spacer region [IGS]), and a state diagnostic laboratory performed the third (universal Mycoplasma [UM]). Overall concordance was good, ranging from 93% to 99%, which was probably a result of low detection rate of M. ovipneumoniae. Due to differences in positive agreement, the quality of concordance between LM40 and both IGS and UM was considered fair. However, the quality of concordance between IGS and UM was excellent. All three PCR methods detected M. ovipneumoniae in a non-Caprinae species (caribou), and the LM40-PCR assay also detected M. ovipneumoniae in additional Caprinae species. The LM40-PCR assay detected M. ovipneumoniae in a larger number of samples than did the other two assays (IGS, UM). Because of potential differences in detection rates, it is critical to consider test parameters when evaluating a host population for the presence of M. ovipneumoniae.