Identification and Characterization of Salmonella Isolates by Automated Ribotyping

Oscar, Thomas P.

doi:10.4315/0362-028x-61.5.519

Cited by 24 publications

(11 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…A growing number of fingerprinting methods have also attempted to identify Salmonella to the serotype level, including randomly amplified polymorphic DNA analysis (51), enterobacterial repetitive intergenic consensus PCR (6), automated ribotyping (38), and PFGE (15). Use of these methods for the identification of Salmonella serotypes, which are based on the establishment and comparison of banding patterns, relies not only on a large and diverse population of samples that reflect the true diversity of all Salmonella serotypes but also on the maintenance and analysis of these patterns in a database, which is not practical.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Multiplex, Bead-Based Suspension Array for Molecular Determination of Common Salmonella Serogroups

et al. 2007

View full text Add to dashboard Cite

We report the development and evaluation of a Salmonella O-group-specific Bio-Plex assay to detect the six most common serogroups in the United States (B, C 1 , C 2 , D, E, and O13) plus serotype Paratyphi A. The assay is based on rfb gene targets directly involved in O-antigen biosynthesis; it can be completed 45 min post-PCR amplification. The assay correctly and specifically identified 362 of 384 (94.3%) isolates tested in comparison to traditional serotyping. Seventeen isolates (4.4%) produced results consistent with what is known about the molecular basis for serotypes but different from the results of traditional serotyping, and five isolates (1.3%) generated false-negative results. Molecular determination of the serogroup for rough isolates was consistent with a common serotype in most instances, indicating that this approach has the potential to provide O-group information for isolates that do not express an O antigen. We also report the sequence of the O-antigenencoding rfb gene cluster from Salmonella enterica serotype Poona (serogroup O13). Compared with other, previously characterized rfb regions, the O13 rfb gene cluster was most closely related to Escherichia coli O127 and O86. The O-group Bio-Plex assay described here provides an easy-to-use, high-throughput system for rapid detection of common Salmonella serogroups.Serotyping of salmonellae is a valuable phenotypic subtyping tool for understanding the epidemiology of this important food-borne pathogen. Salmonella isolates are serotyped using the Kauffmann-White scheme according to their O, H, and Vi antigens (4, 40). The O antigen contains multiple repeats of an oligosaccharide unit (O unit), which, together with lipid A and core oligosaccharides, form the lipopolysaccharide present in the outer membranes of gram-negative bacteria (7). Many of the genes required for O-antigen biosynthesis are organized in a large regulon termed the rfb gene cluster (47). rfb gene clusters have been characterized from a growing number of gram-negative bacteria; this operon is located between galF and gnd in Salmonella enterica and Escherichia coli (49). In general, rfb genes have a low GϩC content (usually less than 40%); the deviation in GϩC content from that of typical S. enterica genes (51%) suggests that rfb DNA originated in species other than S. enterica and was captured by lateral gene transfer (46, 56). Typically, three classes of genes are found in rfb clusters: (i) genes for synthesis of nucleotide sugars specific to the respective O antigen, (ii) sugar transferase genes to build the O subunit, and (iii) the O-antigen polymerase (wzy) and transport protein (wzx) genes for assembly of the O subunit into the O antigen (49).There are 46 O serogroups described in the KauffmannWhite scheme; serogroups were originally designated by alphabetic letters, and then it was necessary to continue with numbers 51 to 67. For consistency in the scheme, all serogroups were given a number designation; however, the most common serogroups (A to E) are commonly designated by ...

show abstract

Section: Discussionmentioning

confidence: 99%

“…Isolates were serotyped according to the method of Brenner and McWhorter-Murlin (4). Serotypes were designated according to the Kauffmann-White scheme (38). Salmonella enterica stock strain 99-0277 (Salmonella enterica serotype Poona, 13,22:z:1,6) was selected for rfb sequence analysis.…”

Section: Methodsmentioning

confidence: 99%

Multiplex, Bead-Based Suspension Array for Molecular Determination of Common Salmonella Serogroups

et al. 2007

View full text Add to dashboard Cite

show abstract

“…With adequate software it becomes possible to compare results from different laboratories. In addition there is the potential for automation of the technique by using the riboprinter (13,25) although this approach would require more flexibility in the use of enzymes and more entries in the database of types. One of the limitations of this technique is the occurrence of both weak and very strong hybridizing bands in the same pattern.…”

Section: Figmentioning

confidence: 99%

Diversity of Strains of Salmonella enterica Serotype Enteritidis from English Poultry Farms Assessed by Multiple Genetic Fingerprinting

et al. 2001

View full text Add to dashboard Cite

Reliable and sufficiently discriminative methods are needed for differentiating individual strains of Salmonella enterica serotype Enteritidis beyond the phenotypic level; however, a consensus has not been reached as to which molecular method is best suited for this purpose. In addition, data are lacking on the molecular fingerprinting of serotype Enteritidis from poultry environments in the United Kingdom. This study evaluated the combined use of classical methods (phage typing) with three well-established molecular methods (ribotyping, macrorestriction analysis of genomic DNA, and plasmid profiling) in the assessment of diversity within 104 isolates of serotype Enteritidis from eight unaffiliated poultry farms in England. The most sensitive technique for identifying polymorphism was PstI-SphI ribotyping, distinguishing a total of 22 patterns, 10 of which were found among phage type 4 isolates. Pulsed-field gel electrophoresis of XbaI-digested genomic DNA segregated the isolates into only six types with minor differences between them. In addition, 14 plasmid profiles were found among this population. When all of the typing methods were combined, 54 types of strains were differentiated, and most of the poultry farms presented a variety of strains, which suggests that serotype Enteritidis organisms representing different genomic groups are circulating in England. In conclusion, geographical and animal origins of Salmonella serotype Enteritidis isolates may have a considerable influence on selecting the best typing strategy for individual programs, and a single method cannot be relied on for discriminating between strains.

show abstract

“…The use of PstI and SphI has been found to give the maximum sensitivity for ribotyping (PS ribotyping) of S. enteritidis strains (Landeras et al, 1996, 1998Liebana et al, 2001). Ribotyping is a technique that requires considerable laboratory expertise, although the method does have some important advantages such as high degree of typeability, stability, good sensitivity, and the possibility of normalisation of data.…”

Section: Discussionmentioning

confidence: 99%

“…With adequate software one can compare results from different laboratories. In addition there is potential for automation of the technique by using the riboprinter (Hilton and Penn, 1998;Oscar, 1998) although this approach would require more flexibility in the use of enzymes, and more entries in the database of types.…”

Section: Discussionmentioning

confidence: 99%

Use of molecular fingerprinting to assist the understanding of the epidemiology of Salmonella contamination within broiler production

Liébana¹,

Crowley²,

Migura-García³

et al. 2002

British Poultry Science

View full text Add to dashboard Cite

1. We analysed Salmonella isolates by conventional sero- and phage-typing, as well as by molecular techniques within the broiler production chain in two integrated companies. The most prevalent serovars were selected for genetic fingerprinting. 2. Isolates were first screened by plasmid profiling; subsequently, the most common plasmid types within the prevalent zoonotic serovars (enteritidis and typhimurium) and S. agama were further characterised by PstI-SphI ribotyping, and XbaI pulsed field gel electrophoresis (PFGE). 3. Salmonella binza, S. kedougou, and S. 4,12:d:- were endemic in the feed mills over long periods of time, and a variety of plasmid types for each of the serovars were found in the premises. 4. A similar situation was found with S. binza and S. senftenberg within the hatchery in company B. The Salmonella serovars which were resident in those locations were also the ones most widely distributed throughout the broiler flocks. 5. Plasmid profiling was useful to subdivide clusters of isolates within serovars, but for each serovar a high percentage (36 to 79%) of the isolates tested fall within a prevalent plasmid type. 6. A more detailed genetic analysis of the isolates by a multiple typing approach allowed for further strain differentiation, and allowed some epidemiological conclusions to be drawn.

show abstract

Identification and Characterization of Salmonella Isolates by Automated Ribotyping

Cited by 24 publications

References 15 publications

Multiplex, Bead-Based Suspension Array for Molecular Determination of Common Salmonella Serogroups

Multiplex, Bead-Based Suspension Array for Molecular Determination of Common Salmonella Serogroups

Diversity of Strains of Salmonella enterica Serotype Enteritidis from English Poultry Farms Assessed by Multiple Genetic Fingerprinting

Use of molecular fingerprinting to assist the understanding of the epidemiology of Salmonella contamination within broiler production

Contact Info

Product

Resources

About