Hybridization among clownfish (genus Amphiprion) is common in the aquarium trade and has also been reported in natural environments. Putative hybrids between Amphiprion chrysopterus and Amphiprion sandaracinos have been identified in a previous genetic study. However, convenient diagnostic tools for hybrid detection for this case are still missing. During the present study, several potential nuclear markers were tested on both parental species and their hybrids collected from Kimbe Bay, Papua New Guinea. Two diagnostic nuclear markers were found that can confidently discriminate the parent species and their hybrids. A haploweb analysis based on those two diagnostic nuclear markers indicated that one backcrossed hybrid was misclassified as one of the purebred parent species. Identification results from these two diagnostic markers were supported by results from microsatellites, mitochondrial gene markers, and morphological traits analysis.