Classification and identification of propionibacteria based on ribosomal RNA genes and PCR

Syst Appl Microbiol. 1998 Jun;21(2):251-9. doi: 10.1016/S0723-2020(98)80030-1.


A rapid method was developed to differentiate the genus Propionibacterium from other genera by using a modified multiplex-PCR (MPCR) approach. Three 16S rRNA-targeted oligonucleotide primers were designed to amplify simultaneously two DNA-fragments in the MPCR assay. The universal primer pair bak11w and bak4 (corresponding to the E. coli 16S rRNA positions 8-25 and 1522-1540, respectively) was used in combination with the primer pair bak4 and gd1 (5'-TGCTTTCGATACGGGTTGAC-3'). The later sequence corresponding to a 16S rRNA motif that is unique for the genus Propionibacterium. Propionibacteria were identified by the amplification of a Propionibacterium-genus specific 900-bp fragment whereas MPCR with DNA from other bacteria generated only a DNA fragment of 1500 bp in amplifications with the two universal primers. The whole procedure including cell lysis, MPCR amplification and analysis can be performed within 1 day, detection limits are at approximately 10(3) cfu propionibacteria (or 35 pg DNA). In addition, the taxonomic situation of the genus Propionibacterium was reexamined using a cycle sequencing strategy. Based on the 16S rDNA, a phylogenetic tree of all the Propionibacterium type strains was reconstructed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Bacterial / chemistry
  • DNA, Ribosomal / chemistry
  • Electrophoresis, Agar Gel
  • Phylogeny*
  • Polymerase Chain Reaction
  • Propionibacterium / chemistry
  • Propionibacterium / classification*
  • Propionibacterium / genetics
  • RNA / chemistry
  • RNA, Bacterial / chemistry
  • RNA, Ribosomal, 16S / chemistry
  • RNA, Ribosomal, 16S / genetics*
  • Sequence Alignment
  • Sequence Analysis, DNA


  • DNA, Bacterial
  • DNA, Ribosomal
  • RNA primers
  • RNA, Bacterial
  • RNA, Ribosomal, 16S
  • RNA

Associated data

  • GENBANK/AJ003055
  • GENBANK/AJ003056
  • GENBANK/AJ003057
  • GENBANK/AJ003058
  • GENBANK/Y10819