DNA fingerprinting is very important for the breeding of new varieties, germplasm resources conservation and management. However, DNA fingerprinting in kenaf using SSR markers is still very limited. In this study, the DNA fingerprint for 96 kenaf accessions, including the control variety Fuhong 952, were constructed on the basis of 144 pairs of SSR primers. The results showed that 144 pairs of primers amplified 375 bands, with the average of 2.6 bands per primer. When genetic similarity coefficient was 0.614, all the kenaf accessions could be divided into two groups, P1 and P2. The group P1 included 52 accessions while P2 had 44 ones. When genetic similarity coefficient was 0.710, all the kenaf accessions could be divided into five subgroups. On the basis of 144 SSRs,85 accessions has specific DNA fingerprints. Among them, HcEMS238 could differate Fuhong 952 from the others. The fingerprint of the other 11 accesssions failed to be constructed due to the presence of high genetic similarity. These results would be helpful in varieties authenticity identification and analysis of genetic diversity in kenaf.