Identification of porcine intestinal spirochetes by PCR-restriction fragment length polymorphism analysis of ribosomal DNA encoding 23S rRNA

D E Barcellos; M de Uzeda; N Ikuta; V R Lunge; A S Fonseca; I I Kader; G E Duhamel

doi:10.1016/s0378-1135(00)00212-1

Identification of porcine intestinal spirochetes by PCR-restriction fragment length polymorphism analysis of ribosomal DNA encoding 23S rRNA

Vet Microbiol. 2000 Jul 31;75(2):189-98. doi: 10.1016/s0378-1135(00)00212-1.

Authors

D E Barcellos¹, M de Uzeda, N Ikuta, V R Lunge, A S Fonseca, I I Kader, G E Duhamel

Affiliation

¹ Instituto de Microbiologia 'Paulo de Góes', Universidade Federal do Rio de Janeiro, R.J., Rio de Janeiro, Brazil.

PMID: 10889409
DOI: 10.1016/s0378-1135(00)00212-1

Abstract

The Brachyspira (formerly Serpulina) species rrl gene encoding 23S ribosomal RNA (rRNA) was used as a target for amplification of a 517bp 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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA Primers / chemistry
DNA, Bacterial / chemistry
DNA, Bacterial / isolation & purification
Deoxyribonucleases, Type II Site-Specific / chemistry
Electrophoresis, Agar Gel / veterinary
Electrophoresis, Polyacrylamide Gel
Intestinal Diseases / diagnosis
Intestinal Diseases / microbiology
Intestinal Diseases / veterinary*
Polymerase Chain Reaction / veterinary
Polymorphism, Restriction Fragment Length
RNA, Ribosomal / chemistry*
RNA, Ribosomal, 23S / chemistry
RNA, Ribosomal, 23S / genetics*
Sequence Alignment
Spirochaetaceae / classification
Spirochaetaceae / genetics
Spirochaetaceae / isolation & purification*
Spirochaetales Infections / diagnosis
Spirochaetales Infections / microbiology
Spirochaetales Infections / veterinary*
Swine
Swine Diseases / diagnosis*
Swine Diseases / microbiology

Substances

DNA Primers
DNA, Bacterial
RNA, Ribosomal
RNA, Ribosomal, 23S
endodeoxyribonuclease AluI
Deoxyribonucleases, Type II Site-Specific
TCGA-specific type II deoxyribonucleases