Use of Random Amplified Polymorphic DNA and Restriction Fragment Length Polymorphism for Typing of Vibrio alginolyticus Isolated from Black Tiger Shrimp Penaeus monodon

作者：Marhual N P^*; Das B K

来源：Journal of Pure and Applied Microbiology, 2009, 3(1): 75-82.

• DNA fingerprinting
• RAPD-PCR
• RFLP
• Vibrio alginolyticus

From 20 isolates representating one strain of Vibrio alginolyticus isolated from Penaeus monodon were selected for the genotyping properties using RAPD-PCR The strains were isolated from different organs as haepatopancreas, external surface, pleopod and telson from prawn and confirmed on Thiosulphate-Citrate Bilesalt Sucrose agar with supplemented with 2% NaCl For the RAPD study, 10 decamer random primers e g OPA, OPB, OPC, OPG, OPH were used for screening Out of 100 primer tested only 34 primers shows amplification of the genomic genomic DNA of V alginolyticus with more than 4 bands Maximum no of bands were produced by OPA 20 i,e 15 DNA finger printing pattern of Valginolyticus showed 3 15 polymorphic bands in 5 series of selected primers Their molecular weight ranging from 0 212 5 238 kbp RFLP analysis has been widely developed as an effective and sensitive method for strain identification Out of 34 primers of RAPD-PCR only 9 primers of OPA (2, 4, 6, 18 and 20) and OPG series (3, 4, 11) shows RFLP product PCR RFLP finger printing pattern of V alginolyticus showed 1 6 polymorphic bands in 2 series (OPA, OPG) of selected primers and their molecular weight ranging from 0 510 3 867 kbp RAPD and RFLP technique appeared to be a reliable and fast technique for genomic study of Vibrio alginolyticus on the basis of their form of isolation and therefore, represents a powerful tool for epidemiological studies of these prawn pathogens

• 出版日期2009-4