Three molecular methods to identify Salmonella enterica serotype Typhimurium DT104: PCR fingerprinting, multiplex PCR and rapid PFGE

Ebner, P

doi:10.1016/s0378-1097(01)00438-4

Cited by 11 publications

(13 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The antigenic differences among the 46 Salmonella O serogroups are due mainly to genetic variation in their respective rfb gene clusters (58). The rfb gene clusters from 15 Salmonella serogroups have been characterized or sequenced: A (32), B (24), C 1 (28), C 2 (5), D 1 (32), D 2 (9), D 3 (9), E (54), O6,14 (18), O17 (19), O18 (19), O35 (56), O30, O50 (50), and O54 (26). The O antigens of serogroups A, B, C 2 , D 1 , D 2 , D 3 , and E have related structures, and their rfb regions are related.…”

mentioning

confidence: 99%

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

mentioning

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

“…Many methods have been developed to phenotypically distinguish between S. enterica serovar Typhimurium isolates, including antibiotic susceptibility profiling, phage typing (1), pulsed-field gel electrophoresis (PFGE) (7), and plasmid profiling (26) as well as various PCR-based techniques (8,15,19).…”

mentioning

confidence: 99%

Prophage Sequences Defining Hot Spots of Genome Variation inSalmonella entericaSerovar Typhimurium Can Be Used To Discriminate between Field Isolates

et al. 2007

View full text Add to dashboard Cite

Sixty-one Salmonella enterica serovar Typhimurium isolates of animal and human origin, matched by phage type, antimicrobial resistance pattern, and place of isolation, were analyzed by microbiological and molecular techniques, including pulsed-field gel electrophoresis (PFGE) and plasmid profiling. PFGE identified 10 profiles that clustered by phage type and antibiotic resistance pattern with human and animal isolates distributed among different PFGE profiles. Genomic DNA was purified from 23 representative strains and hybridized to the composite Salmonella DNA microarray, and specific genomic regions that exhibited significant variation between isolates were identified. Bioinformatic analysis showed that variable regions of DNA were associated with prophage-like elements. Subsequently, simple multiplex PCR assays were designed on the basis of these variable regions that could be used to discriminate between S. enterica serovar Typhimurium isolates from the same geographical region. These multiplex PCR assays, based on prophage-like elements and Salmonella genomic island 1, provide a simple method for identifying new variants of S. enterica serovar Typhimurium in the field.

show abstract

“…By use of 37 different phages, serotype Typhimurium can be divided in more than 210 phage types (1). Besides serotyping and phage typing, powerful bacterial molecular typing methods, such as plasmid profiling, pulsed-field gel electrophoresis (PFGE), IS200 typing, ribotyping, random amplified polymorphic DNA analysis, and amplified fragment length polymorphism, are used for epidemiological investigation of salmonellae (3,7,8,13,16). These techniques are useful for defining clonal relationships between strains (17) and for assessing the distribution of Salmonella strains within food-processing environments (10,15).…”

mentioning

confidence: 99%

Phenotypic and Molecular Typing of Salmonella Strains Reveals Different Contamination Sources in Two Commercial Pig Slaughterhouses

Botteldoorn¹,

Herman²,

Rijpens³

et al. 2004

Appl Environ Microbiol

View full text Add to dashboard Cite

This study aimed to define the origin of Salmonella contamination on swine carcasses and the distribution of Salmonella serotypes in two commercial slaughterhouses during normal activity. Salmonellae were isolated from carcasses, from colons and mesenteric lymph nodes of individual pigs, and from the slaughterhouse environment. All strains were serotyped; Salmonella enterica serotype Typhimurium and Salmonella enterica serotype Derby isolates were additionally typed beyond the serotype level by pulsed-field gel electrophoresis (PFGE) and antibiotic resistance profiling (ARP); and a subset of 31 serotype Typhimurium strains were additionally phage typed. PFGE and ARP had the same discriminative possibility. Phage typing in combination with PFGE could give extra information for some strains. In one slaughterhouse, 21% of the carcasses were contaminated, reflecting a correlation with the delivery of infected pigs. Carcass contamination did not result only from infection of the corresponding pig; only 25% of the positive carcasses were contaminated with the same serotype or genotype found in the corresponding feces or mesenteric lymph nodes. In the other slaughterhouse, 70% of the carcasses were contaminated, and only in 4% was the same genotype or serotype detected as in the feces of the corresponding pigs. The other positive carcasses in both slaughterhouses were contaminated by genotypes present in the feces or lymph nodes of pigs slaughtered earlier that day or from dispersed sources in the environment. In slaughterhouses, complex contamination cycles may be present, resulting in the isolation of many different genotypes circulating in the environment due to the supply of positive animals and in the contamination of carcasses, probably through aerosols.

show abstract

Three molecular methods to identify Salmonella enterica serotype Typhimurium DT104: PCR fingerprinting, multiplex PCR and rapid PFGE

Cited by 11 publications

References 7 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

Prophage Sequences Defining Hot Spots of Genome Variation inSalmonella entericaSerovar Typhimurium Can Be Used To Discriminate between Field Isolates

Phenotypic and Molecular Typing of Salmonella Strains Reveals Different Contamination Sources in Two Commercial Pig Slaughterhouses

Contact Info

Product

Resources

About