16S to 23S rRNA spacer fragment length polymorphism of Salmonella enterica at subspecies and serotype levels

Abstract

. 2000. The variation in the lengths of the internal transcribed spacer (ITS) between 16S and 23S rRNA genes of 101 strains representing 58 serotypes of Salmonella enterica (used for Salm. choleraesuis) subsp. enterica (I), salamae (II), arizonae (IIIa), diarizonae (IIIb), houtenae (IV) and indica (VI) was used for typing by PCR ampli®cation. Ten fragment lengths were observed by denaturing polyacrylamide gel electrophoresis on an automatic DNA sequencer resulting in 21 unique fragment patterns. Ten out of the 58 serotypes showed speci®c patterns but 48 serotypes were not fully differentiated. More than one ITS pattern was observed in seven serotypes. Five of the 21 fragment patterns contained representatives of more than one subspecies. Under non-denaturing electrophoresis conditions, serotype speci®city was obtained but precise ITS fragment length determination was not possible. DNA sequence comparison between ITSs of individual rrn operons is needed to further interpret ITS diversity within Salm. enterica at serotype and subspecies levels.

Molecular epidemiology and population genetics of Salmonella subspecies diarizonae in sheep in Norway and Sweden

“…The two serovars that carried similar antigen determinants interspaced between S. IIIb 61:k:1,5, 7isolates, while S. IIIb 48:i:z, without any common antigen determinants was the least-related strain. This inconsistent correlation between serovars and genotype is in accord with a recently published paper where 48 out of 58 serovars, representing all six subspecies of Salmonella enterica, were not fully differentiated, and more than one genotype was observed in seven serovars [30]. Such lack of correlation has also been described from genetic investigations of serovars in subspecies enterica (I) [24].…”

Genetic relationships among avian isolates classified as Pasteurella haemolytica, ‘Actinobacillus salpingitidis’ or Pasteurella anatis with proposal of Gallibacterium anatis gen. nov., comb. nov. and description of additional genomospecies within Gallibacterium gen. nov.

“…The primer for reverse amplification was located on the rrl gene with similarity to the E. coli rrnB positions 115-130 (23S-130, 59-GGGTTTCCCCATTCGG) (Christensen et al, 2000b). The primers for forward amplification of the ITS region were located on the rrs gene at E. coli rrnB positions 1525-1541 (16S-1525, 59-GGTTGGATCACCTCCTT) (Christensen et al, 2000b), 1485-1504 (16S1, 59-TGGGGTGAAG-TCGTAACAAG; Privitera et al, 1998) and 1337-1353Fussing et al, 1998). Fragments generated with the three primer sets are 149, 189, 337 bp longer than the 16S-23S ITS.…”

Phenotypic and Genotypic Eligible Methods for Salmonella Typhimurium Source Tracking

“…After gel electrophoresis and transference of the fragments to a membrane, they are probed with a region of the rRNA operon to reveal the patterns of rRNA genes (Bailey et al, 2002 ). The sequence variations in the flanking restriction sites result in a small number of different RFLP banding profiles of the conserved domains of the 16S and 23S rRNA genes that are quite simple to interpret (Christensen et al, 2000 ). However, due to complexity and technicality, manual ribotyping is not a method of choice to typing pathogens (Tenover et al, 1997 ).…”