The Javan tiger Panthera tigris sondaica and the Bali tiger P. tigris balica were categorized as Extinct on the IUCN Red List in both 2003 and 2008, leaving only the Sumatran subspecies P. tigris sumatrae extant in Indonesia. There have, however, been occasional, more recent reports of the Javan tiger but without conclusive evidence. Here, a potential observation in 2019 of a Javan tiger in a community plantation near the village of Cipendeuy in the forest of South Sukabumi, West Java, and a single hair found on a fence nearby, are assessed. The cytochrome b mitochondrial DNA (mtDNA) gene sequence of the putative Javan tiger hair were compared with that of a Javan tiger specimen in Museum Zoologicum Bogoriense, collected in 1930; hair samples of several tiger subspecies and the Javan leopard Panthera pardus melas were used as controls. The results showed that the genetic distances (d) of the putative Javan tiger hair with the Sumatran, Bengal P. tigris tigris and Amur P. tigris altaica tigers and the Javan leopard are 0.074 ± SE 0.009, 0.071 ± SE 0.009, 0.072 ± SE 0.009 and 0.088 ± SE 0.010, respectively, whereas the genetic distance of the putative Javan tiger hair with the Javan tiger museum specimen is 0.040 ± SE 0.006. In addition, phylogenetic trees showed that the putative Javan tiger hair sample belongs to the same group as the museum specimen of the Javan tiger, but is differentiated from other tiger subspecies and the Javan leopard. Whether the Javan tiger still occurs in the wild needs to be confirmed with further genetic and field studies.