Spodoptera litura F. is a new pest that potentially becomes a serious pest threatening palm oil industry. The control can be carried out using biological control agents S.litura Nucleopolyhedrovirus (SlNPV). Molecular approach using polymerase chain reaction (PCR) has been used to characterize the SlNPV isolate. The aim of this study was to determine the molecular characteristics of SlNPV Labuhan Batu isolate based on DNA sequence analysis. The NPV isolate was collected from oil palm plantation Negeri Lama Estate Group, PT. Hari sawit Jaya, Asian Agri, Labuhan Batu. The SlNPV DNA extraction was conducted using the DNA gSYNC kit. The amplification of SlNPV DNA was done by specific primers with oligonucleotide sequences forward 5′-ACGTTACGATCTAGATCC-3′ and reverse 5′-AACGTTAGACAGACATAT-3′ with ±700bp of amplicon. Homology and phylogeny characteristics were used to determine the relationship of SlNPV isolate from Labuhan Batu to NPV from other countries were reported in the GeneBank of National Center for Biotechnology Information (NCBI). The results showed that SlNPV isolate from Labuhan Batu had the closest related to SlNPV from Australia, SliNPV from Canada and SlNPV from China, with the similarity percentage 100%, 96% and 90% respectively. They belong to the same group as the NPVs that infected the Spodoptera genus.