J Vet Sci.  2006 Sep;7(3):277-280. 10.4142/jvs.2006.7.3.277.

The 23S rRNA gene PCR-RFLP used for characterization of porcine intestinal spirochete isolates

• Kim TJ
• Lee JI

• KMID: 1089913
• DOI: http://doi.org/10.4142/jvs.2006.7.3.277

Abstract

Using three reference strains of Brachyspira hyodysenteriae (B204, B234, B169), one B. pilosicoli (P43/6/78), one B. murdochii (56-150), one B. intermedia (PWS/A), one B. innocens (B256) and ten Korean isolates, PCR-RFLP analysis of DNA encoding 23S rRNA was performed to establish a rapid and accurate method for characterizing porcine intestinal spirochetes. Consequently, B. hyodysenteriae and B. pilosicoli revealed different restriction patterns; however, the other three species shared the same pattern. These findings are not consistent with a prior report. Differences in 23S rRNA gene sequences, between two B. murdochii strains, 56-150 and 155-20, were observed. These results indicate that 23S rRNA PCR-RFLP could be used as an identification method for pathogenic Brachyspira spp. (B. hyodysenteriae and B. pilosicoli) as well as an epidemiological tool for characterizing spirochetes isolated from swine.

Keyword

Brachyspira spp.; PCR-RFLP; 23S rRNA; spirochete

MeSH Terms

Animals
DNA, Bacterial/genetics
Dysentery, Bacillary/diagnosis/microbiology/*veterinary
Korea
Phylogeny
Polymerase Chain Reaction/methods/*veterinary
Polymorphism, Restriction Fragment Length
RNA, Ribosomal, 23S/chemistry/*genetics
Spirochaetales/*genetics/*isolation&purification
Spirochaetales Infections/diagnosis/microbiology/*veterinary
Swine
Swine Diseases/diagnosis/*microbiology

Figure

• 

  Fig. 1 PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with TaqI. Lane M: 100 bp DNA ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.

• 

  Fig. 2 PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with AluI. Lane M: 100 bp DNA ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.

• 

  Fig. 3 Phylogenic analyses of 23S rRNA gene sequences in Brachyspira spp. The genetic distances are proportional to the relative sequence deviations between individual nucleotide sequences.

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