Repetitive sequence based polymerase chain reaction to differentiate close bacteria strains in acidic sites

To study the diversity of bacteria strains newly isolated from several acid mine drainage(AMD) sites in China, repetitive sequence based polymerase chain reaction (rep-PCR), a well established technology for diversity analysis of closely related bacteria strains, was conducted on 30 strains of bacteria Leptospirillum ferriphilium, 8 strains of bacteria Acidithiobacillus ferrooxidans, as well as the Acidithiobacillus ferrooxidans type strain ATCC (American Type Culture Collection) 23270. The results showed that, using ERIC and BOX primer sets, rep-PCR produced highly discriminatory banding patterns. Phylogenetic analysis based on ERIC-PCR banding types was made and the results indicated that rep-PCR could be used as a rapid and highly discriminatory screening technique in studying bacterial diversity, especially in differentiating bacteria within one species in AMD.