Identification of porcine intestinal spirochetes by PCR-restriction fragment length polymorphism analysis of ribosomal DNA encoding 23S rRNA
The Brachyspira (formerly Serpulina) species rrl gene encoding 23S ribosomal RNA (rRNA) was used as a target for amplification of a 517 bp DNA fragment by polymerase chain reaction (PCR). The primers for PCR amplification had sequences that were conserved among Brachyspira 23S rRNA gene and were des...
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Veröffentlicht in: | Veterinary microbiology 2000-07, Vol.75 (2), p.189-198 |
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Sprache: | eng |
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Zusammenfassung: | The
Brachyspira (formerly
Serpulina) species
rrl gene encoding 23S ribosomal RNA (rRNA) was used as a target for amplification of a 517
bp DNA fragment by polymerase chain reaction (PCR). The primers for PCR amplification had sequences that were conserved among
Brachyspira 23S rRNA gene and were designed from nucleotide sequences of
Brachyspira hyodysenteriae,
Serpulina intermedia,
Brachyspira innocens and
Brachyspira pilosicoli available from the GenBank database. Digestion of PCR-generated products from reference and field isolates of swine intestinal spirochetes with restriction enzymes
Taq I and
Alu I revealed five restriction fragment length polymorphism (RFLP) patterns. Each RFLP pattern corresponded to previously established genetic groups including
B. hyodysenteriae (I),
S. intermedia/
B. innocens (II),
Brachyspira murdochii (III),
B. pilosicoli (IV) and
B. alvinipulli (V). The 23S rRNA PCR/RFLP provided a relatively simple genotypic method for identification of porcine pathogenic
B. hyodysenteriae and
B. pilosicoli. |
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ISSN: | 0378-1135 1873-2542 |
DOI: | 10.1016/S0378-1135(00)00212-1 |