Molecular fingerprinting of pathogenic Enterococcus isolates using Random Amplified Polymorphic DNA Analysis
By: Ragaa A. Eissa, Mohamed M. Hassan, Ahmed Gaber, Mohamed A. El-Awady
Key Words: RAPD-PCR, fingerprinting, Taif, Pathogenic Enterococcus.
J. Bio. Env. Sci. 8(2), 208-215, February 2016.
Enterococcus sp. is considered to be one of the most prevalent species in the oral cavity, particularly in endodontic infections. The aim of the present study was to investigate the molecular fingerprinting of Enterococcus sp in patient’s urine diversity by randomly amplified polymorphic DNA (RAPD-PCR) analysis of Enterococcus. Specific results for fingerprinting were obtained by the five primers retained for RAPD analysis produced different fragment patterns with varied number of bands. The primers yielded a total of 71 distinct bands 67.6 % were considered as polymorphic bands and 32.4% were considered as monomorphic bands. The OPA-01 primer has showed the highest polymorphism 76.9 % and total of 15 bands ranged from 300 bp-1700 bp. While, the OPD-05 primer has showed the lowest polymorphism 57.2 % and total of 14 bands ranged from 190 bp-2200 bp. Interestingly, OPA-01 primer was a unique negative marker for isolate 7 and, on the other hand, gives a positive one unique band only with isolate 9. The Dendrogram based on RAPD results grouped the thirteen Enterococcus isolates into two different clusters with about 82.2% genetic similarity. Moreover, the genetic distance among Enterococcus isolates was relatively low. The smallest genetic distance (0.167) was estimated between E 10 and E 13. Results indicate the presence of sufficient nuclear DNA level variations among studied Enterococcus isolates using RAPD markers, and the RAPD data might be a good source of information about the diversity of native Enterococcus isolates.