Comparison of pulsed field gel electrophoresis and repetitive sequence polymerase chain reaction as genotyping methods for detection of genetic diversity and inferring transmission of Salmonella

Weigel, Ronald M.; Qiao, Baozhen; Teferedegne, B.; Suh, Dong Kyun; Barber, D. A.; Isaacson, Richard E.; White, Bryan A.

doi:10.1016/j.vetmic.2004.02.009

Cited by 77 publications

(56 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Apesar da grande homologia das seqüências rEP e ErIc entre as enterobactérias, DIMrI et al [12] observaram uma variação em quantidade e localização destes elementos no genoma de E. coli, Shigella, Salmonella, Klebsiella, Enterobacter, Serratia, Erwinia e Proteus. A rEP e ErIc são métodos simples, rápidos e sensíveis que podem ser utilizados para discriminar sorovares relacionados [4,20,31,34].…”

Section: Discriminação De Sorovares De Salmonella Spp Isolados De Caunclassified

“…Muitos autores têm mostrado que tanto a rEP como a ErIcPcr são métodos genéticos sorovar específico para o gênero Salmonella [4,20,30,34]. A característica sorovar específica da rEP-Pcr para esse gênero foi relatada por BENNASAr et al [3], embora utilizando temperatura de anelamento inferior (33ºc) à utilizada no presente trabalho (45ºc).…”

Section: -Discussãounclassified

“…WEIGEl et al [34] mostraram que a eletroforese em gel de pulso alternado (PFGE) é tão discriminativa como a rEP-Pcr para o gênero Salmonella. Por outro lado, DESAI, THrElFAll & STANlEY [11] obtiveram bons resultados ao utilizarem a fluorescent amplified-fragment legth polymorphism (FAFLP) para subtipificar Salmonella Enteriditis fagotipo 4. recentemente, FErNANDES et al [15] mostraram que a ribotipagem foi capaz de diferenciar seis ribotipos de Enteriditis no fagotipo 8 e três no fagotipo 4.…”

Section: -Discussãounclassified

See 2 more Smart Citations

Discriminação de sorovares de Salmonella spp. isolados de carcaças de frango por REP e ERIC-PCR e fagotipagem do sorovar Enteriditis

Alcocer¹,

Oliveira²,

Vidotto³

et al. 2006

Ciênc. Tecnol. Aliment.

View full text Add to dashboard Cite

RESUMOSalmonelose é a infecção bacteriana de origem alimentar mais freqüente no Paraná, Brasil, e os surtos estão associados, principalmente, ao consumo de ovos, carne de aves e derivados. Os objetivos deste trabalho foram identificar os sorovares de Salmonella isolados de carcaças de frango e caracterizá-los molecularmente por rEP e ErIc-Pcr, assim como identificar os fagotipos de Salmonella Enteriditis. Dos 25 isolados de Salmonella spp. analisados, 18 foram identificados como Enteriditis, 4 como Braenderup, 2 como Worthington e 1 como infantis. Dos 18 isolados de Enteriditis, 14 foram PT4, 2 PT4a, 1 PT7 e 1 rDNc, por se tratar de colônia rugosa. rEP-Pcr forneceu padrão eletroforético distinto de 10 a 13 bandas distribuídas entre 120 e 2072 pb para cada sorovar diferente testado. A ErIc-Pcr mostrou um padrão de 4 a 5 bandas entre 180 e 1000 pb e foi menos discriminativa quando comparada à rEP-Pcr. Os resultados encontrados confirmaram que a fagotipagem é uma ferramenta útil e discriminativa para o sorovar Enteriditis. Apesar do pequeno número de sorovares testados, os resultados sugerem que a rEP-Pcr parece ser um método atrativo a ser utilizado no futuro para a discriminação preliminar de sorovares de Salmonella. Palavras-chaves: Salmonella Enteriditis, fagotipos, sorovar, rEP-Pcr, ErIc-Pcr. SUMMARYDIScrIMINATION OF Salmonella SErOVArS ISOlATED FrOM cHIcKEN MEAT BY rEP AND ErIc-Pcr AND PHAGOTYPING OF Enteriditis SOrOVAr. Salmonellosis is the most prevalent bacterial food-borne disease in the State of Paraná, Brazil, and the outbreaks are often associated with consumption of poultry products. The aim of this study was to serotype Salmonella strains isolated from chicken carcasses and characterize them molecularly using rEP and ErIc-Pcr. The phage types of Salmonella Enteriditis were also identified. Of the 25 Salmonella strains analysed, 18 were identified as Enteriditis, 4 as Braenderup, 2 as Worthington and 1 as infantis. Of the 18 Enteriditis isolates, 14 were PT4, 2 PT4a, 1 PT7 and 1 "reacted, but did not conform" -rDNc. Distinct rEP-Pcr profiles with 10 to 13 fragments distributed between 120 and 2072 pb were easily obtained for each serovar tested. ErIc-Pcr showed patterns of 4 to 5 fragments between 180 and 1000 pb and less discriminatory power than rEPPcr. The results confirmed that phage typing is a useful tool to differentiate into sorovar Enteriditis. Although it is recognized that only a limited number of strains was tested in this study, the results suggest that rEP-Pcr could be an attractive choice to be used in the future as a preliminary method of Salmonella sorovar discrimination.

show abstract

Section: Discriminação De Sorovares De Salmonella Spp Isolados De Caunclassified

Section: -Discussãounclassified

See 1 more Smart Citation

Discriminação de sorovares de Salmonella spp. isolados de carcaças de frango por REP e ERIC-PCR e fagotipagem do sorovar Enteriditis

Alcocer¹,

Oliveira²,

Vidotto³

et al. 2006

Ciênc. Tecnol. Aliment.

View full text Add to dashboard Cite

show abstract

“…Phenotypic methods play an important role in identification of genus level. Serotyping, based on the Kauffmann-White scheme, remains the standard for classification of Salmonella isolates in outbreak investigations but now has been supplemented by a range of molecular genotyping methods [1,2]. In recent years, molecular-based techniques, such as PulsedField Gel Electrophoresis (PFGE), Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD), Automatic Ribo-typing, rep-PCR, Multi-Locus Sequence Typing (MLST) have been shown to be useful methods for discrimination among isolates of pathogens [3,4].…”

Section: Introductionmentioning

confidence: 99%

RAPD-PCR as a Molecular Discriminative Technique for Human Pathogenic Bacteria - A Review

Pal¹

2015

ILNS

View full text Add to dashboard Cite

ABSTRACT. The application of random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) was found to be a simple, cheap and rapid tool to discriminate human pathogenic bacterial isolates especially at intraspecific level. This molecular biological technique relies on the use of random oligonucleotide primers that arbitrarily amplifies specific regions of the genome which gives rise to a unique genomic fingerprint of the strains under investigations. With continued development of novel molecular-based technologies for rapid, high-throughput detection of food borne pathogenic bacteria, the future of conventional microbiological methods such as viable cell enumeration, selective isolation of bacteria on commercial media, and immunoassays seems tenuous. Approaches that enhance recovery of sub lethally injured bacteria, differentiation among species, differentiation among bacteria of interest using biochemical profiling, enumeration using impedance technology, techniques to confirm the presence of target pathogens using immunological methods, and bioluminescence applications for hygiene monitoring are of utmost need in identifying and combating the human pathogenic isolates. The aim of this study is to estimate the efficiency of RAPD-PCR technique in assessing the genetic diversity of diseases causing bacterial isolates. The use of RAPD-PCR in evaluating the genomic variability among the pathogenic strains belonging to different genus are also been discussed in the present report.

show abstract

“…The gold standard for differentiating epidemiologically related strains of S. enterica is the analysis of polymorphisms through pulsed-field gel electrophoresis (PFGE) [4]. However, analysis of repetitive DNA elements has also been proven to be successful [5,6], and even equally or more effective than PFGE for that purpose [7,8]. Additionally, it has been reported that random amplified polymorphic DNA (RAPD) assays combined with other typing techniques are useful for monitoring outbreaks and clinical cases of gastroenteritis caused by serotypes of S. enterica [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Investigation of a food-borne Salmonella Oranienburg outbreak in a Mexican prison

Vázquez-Garcidueñas

Romero-Pérez

Figueroa-Aguilar

et al. 2014

J Infect Dev Ctries

View full text Add to dashboard Cite

Introduction: Gastroenteritis outbreaks in prisons represent a public health risk worldwide. Identifying and characterizing the etiological agents of gastroenteritis outbreaks in prisons is important for implementing effective prevention and infection control measures. We present the first studied case of a gastroenteritis outbreak in a Mexican prison. Methodology: Rectal swab samples were obtained from affected inmates. Standard microbiological techniques were used for isolating Salmonella enterica. Isolates were typed by PCR assays of DNA repetitive elements (ERIC, BOX, REP) and RAPD. Antibiotic resistance profiles were performed by the Kirby-Bauer method. Results: S. enterica serotype Oranienburg was responsible for the outbreak affecting 150 inmates. All patients presented diarrhea, and 70% of them also presented vomiting, with no fatal cases. The origin of the outbreak was undetermined due to the difficulty of gathering epidemiological information, but was likely the result of consumption of shrimp broth or a cantaloupe melon beverage. REP, BOX, and ERIC analyses of 26 serotype Oranienburg strains resulted in Simpson discrimination index (D) values of 0, 0.5507, and 0.5661, respectively. The D values from DG93-RAPD analyses and from the combined ERIC-BOX-DG93 markers were 0.7753 and 0.6092, respectively. All strains showed multiresistance to antibiotics. Conclusions: This is the only studied case of a gastroenteritis outbreak in a Mexican prison, and of the first such outbreak caused by serotype Oranienburg. The combined ERIC, BOX, and RAPD markers adequately assessed the genotype diversity of analyzed strains. Penitentiary personnel or inmates involved in outbreaks might spread multiresistant strains outside of the facility.

show abstract

Comparison of pulsed field gel electrophoresis and repetitive sequence polymerase chain reaction as genotyping methods for detection of genetic diversity and inferring transmission of Salmonella

Cited by 77 publications

References 35 publications

Discriminação de sorovares de Salmonella spp. isolados de carcaças de frango por REP e ERIC-PCR e fagotipagem do sorovar Enteriditis

Discriminação de sorovares de Salmonella spp. isolados de carcaças de frango por REP e ERIC-PCR e fagotipagem do sorovar Enteriditis

RAPD-PCR as a Molecular Discriminative Technique for Human Pathogenic Bacteria - A Review

Investigation of a food-borne Salmonella Oranienburg outbreak in a Mexican prison

Contact Info

Product

Resources

About