An undescribed relapsing fever group Borrelia species was detected in a male Haemaphysalis semermis tick infesting a rural cat in an indigenous population in Pahang National Park, Peninsular Malaysia. The 16 S rRNA gene sequence revealed close similarity of this variant to several undescribed Borrelia species and Borrelia theileri, with genetic distances ranging from 0.58 to 0.72%. Furthermore, the flaB, gyrB, and the concatenated 16 S rRNA + flaB + gyrB sequence analyses demonstrated that this variant is distinctly separated from multiple undescribed Borrelia species, Borrelia miyamotoi, and B. theileri, with genetic distances ranging from 3.41 to 7.00%. This study not only reports the first Borrelia found in H. semermis but also suggests that it forms a distinct clade within the relapsing fever group in Peninsular Malaysia.