The dermatoglyphic characters can measure genetics' variations within and between human populations. This study aims to characterize the fingerprints traits in the eastern part of Libya, to analyze bilateral and bisexual differences within Libyan sub-populations. The study conducted on 1101 subjects in six cities; Benghazi, Al-Bayda, Al-Marj, Dernah, and Jalu-Ojla. Data included age, sex, and the parameters of digital dermatoglyphics have been recorded and analyzed. For statistical analysis, the frequency distributions for digital patterns were tested by chi-square, ridge were tested by student t test, and genetics distance were estimated by using the phylogenetic analysis. Total finger ridge count was significantly (p<0.01) higher in males (138.35) than females (131.46). Dindices were highest in Jalu-Ojla, Al-Bayda, Benghazi, Dernah and Al-Marj, respectively. F-indices were highest in Jalu-Ojla, Dernah, Benghazi Al-Bayda and Al-Marj, respectively; Total ridge counts were generally higher in males than females except in Jalu-Ojla and Al-Marj cities, where the latter two cities were equal. Arches and radial loops, ulnar loops were generally higher than the Whorls that were found in lower level due to the influence of consanguinity marriages, founder effect and random drift of genes. Total ridge counts were higher in males than females and in the right-hand, but not in left-hand for all the study areas. Therefore, the gender, bilateral side of the hand and the consanguinity level within the Libyan sub-population should be considered when establishing dermatoglyphic pattern indices for the Libyan population.