Identification of Salmonella spp. isolates from chicken abattoirs by multiplex-PCR

Cortez, A.L.L.; Carvalho, A. C. F. B.; Ikuno, A. A.; Bürger, Karina Paes; Vidal, Ana Maria Centola

doi:10.1016/j.rvsc.2006.03.006

Cited by 60 publications

(41 citation statements)

References 14 publications

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“…521 bp [41] All identified isolates were subjected to an analysis of PCR-RFLP of the 16S rRNA genes, while several restriction endonucleases were separately used to digest the PCR products. The genotyping pattern of strains with enzymes including AluI, BglI, HindIII, EcoRI and BgIII (Thermo Scientific, USA) demonstrated that they can be classified into two main groups.…”

Section: Resultsmentioning

confidence: 99%

Genetic diversity of food originated Salmonella isolates

Karatuğ

Yüksel

Akçelik

et al. 2018

Biotechnology & Biotechnological Equipment

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Currently, Salmonella enterica is the most common bacterial foodborne pathogen, causing serious extraintestinal disease. Typing methods play an important role on pathogens' source tracking, knowing the source(s) of bacteria in pharmaceutical sciences, preventing and controlling the diarrhea and food-poisoning outbreaks. The purpose of this study is to use different moleculer typing methods to determine the genetic variability of 38 foodborne Salmonella isolates that were previously identified by biochemical tests. The methods were evaluated by four molecular techniques including 16S rRNA sequencing, PFGE, PCR-RFLP and invA-spvC PCR. 16S rRNA sequencing results showed that four of the 38 isolates were Escherichia coli, Proteus mirabilis and Citrobacter murliniae, and the others were Salmonella enterica. Thirty-eight strains were subtyped by XbaI-PFGE into 20 profiles with different clusters, while they were subtyped by 16S rRNA-RFLP into 9 profiles with a single cluster. Out of two Salmonella isolates, the invasion gene (invA) was detected in all other Salmonella isolates (94%) and the virulence gene (spvC) was detected in 11% of Salmonella isolates. Our results suggested that the PFGE subtyping is the prominent method for the evaluation and benchmarking of molecular subtyping.

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Section: Resultsmentioning

confidence: 99%

Genetic diversity of food originated Salmonella isolates

Karatuğ

Yüksel

Akçelik

et al. 2018

Biotechnology & Biotechnological Equipment

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“…Differentiation of S. Typhimurium and S. Enteritidis could be made based on the presence in the former and absence in the latter of a restriction site for the enzyme Kpnl in the pefA gene [16]. To differentiate S. Typhimurium and S. Enteritidis, we have used PCR with a primer set targeting Kpnl enzyme target sequence in pefA gene.…”

Section: Discussionmentioning

confidence: 99%

“…A multiplex PCR method developed by [16] was used, with modifications in the primer sets. The sets of primers used in the multiplex PCR assay were Pef A-Fwd-5 0 -TTC CATTATTGCACTGGGTG-3 0 , PefA-Rev-5 0 -AAGCCACTGCGAAAGATGCC-3 0 , that were selected based on the 5 0 -3 0 conserved region of the fimbrial virulence gene (pefA).…”

Section: Molecular Characterizationmentioning

confidence: 99%

Molecular Epidemiology of Nontyphoidal Salmonella in Poultry and Poultry Products in India: Implications for Human Health

et al. 2015

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Human infections with non-typhoidal Salmonella (NTS) serovars are increasingly becoming a threat to human health globally. While all motile Salmonellae have zoonotic potential, Salmonella Enteritidis and Salmonella Typhimurium are most commonly associated with human disease, for which poultry are a major source. Despite the increasing number of human NTS infections, the epidemiology of NTS in poultry in India has not been fully understood. Hence, as a first step, we carried out epidemiological analysis to establish the incidence of NTS in poultry to evaluate the risk to human health. A total of 1215 samples (including poultry meat, tissues, egg and environmental samples) were collected from 154 commercial layer farms from southern India and screened for NTS. Following identification by cultural and biochemical methods, Salmonella isolates were further characterized by multiplex PCR, allele-specific PCR, enterobacterial repetitive intergenic consensus (ERIC) PCR and pulse field gel electrophoresis (PFGE). In the present study, 21/1215 (1.73 %) samples tested positive for NTS. We found 12/392 (3.06 %) of tissue samples, 7/460 (1.52 %) of poultry products, and 2/363 (0.55 %) of environmental samples tested positive for NTS. All the Salmonella isolates were resistant to oxytetracycline, which is routinely used as poultry feed additive. The multiplex PCR results allowed 16/21 isolates to be classified as S. Typhimurium, and five isolates as S. Enteritidis. Of the five S. Enteritidis isolates, four were identified as group D Salmonella by allele-specific PCR. All of the isolates produced different banding patterns in ERIC PCR. Of the thirteen macro restriction profiles (MRPs) obtained by PFGE, MRP 6 was predominant which included 6 (21 %) isolates. In conclusion, the findings of the study revealed higher incidence of contamination of NTS Salmonella in poultry tissue and animal protein sources used for poultry. The results of the study warrants further investigation on different type of animal feed sources, food market chains, processing plants, live bird markets etc., to evaluate the risk factors, transmission and effective control measures of human Salmonella infection from poultry products.

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“…This gene is highly conserved in almost all Salmonella serotypes (7,17,44), and detection of Salmonella based on the presence of this gene has previously been reported (11,12,24). invA mRNA has been used as a biomarker for active cells (13,18,26); it has been discussed, however, whether transcription of the invA gene may differ with different physiological states of the cell, which may affect assay specificity (13).…”

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confidence: 95%

Influence of Temperature and Predation on Survival of Salmonella enterica Serovar Typhimurium and Expression of invA in Soil and Manure-Amended Soil

Bælum

Fredslund

et al. 2010

Appl Environ Microbiol

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The effects of three temperatures (5, 15, and 25°C) on the survival of Salmonella enterica serovar Typhimurium in topsoil were investigated in small microcosms by three different techniques: plate counting, invA gene quantification, and invA mRNA quantification. Differences in survival were related to the effect of protozoan predation. Tetracycline-resistant Salmonella serovar Typhimurium was inoculated into soil and manure-amended soil at 1.5 ؋ 10 8 cells g soil ؊1 . Population densities were determined by plate counting and by molecular methods and monitored for 42 days. Simultaneous extraction of RNA and DNA, followed by quantitative PCR, was used to investigate invA gene levels and expression. Analysis by these three techniques showed that Salmonella serovar Typhimurium survived better at 5°C. Comparing DNA and CFU levels, significantly higher values were determined by DNA-based techniques. invA mRNA levels showed a fast decrease in activity, with no detectable mRNA after an incubation period of less than 4 days in any of the soil scenarios. A negative correlation was found between Salmonella serovar Typhimurium CFU levels and protozoan most probable numbers, and we propose the role of the predator-prey interaction as a factor to explain the die-off of the introduced strain by both culture-and DNA quantification-based methods. The results indicate that temperature, manure, and protozoan predation are important factors influencing the survival of Salmonella serovar Typhimurium in soil.

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Identification of Salmonella spp. isolates from chicken abattoirs by multiplex-PCR

Cited by 60 publications

References 14 publications

Genetic diversity of food originated Salmonella isolates

Genetic diversity of food originated Salmonella isolates

Molecular Epidemiology of Nontyphoidal Salmonella in Poultry and Poultry Products in India: Implications for Human Health

Influence of Temperature and Predation on Survival of Salmonella enterica Serovar Typhimurium and Expression of invA in Soil and Manure-Amended Soil

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