Use of Polymerase Chain Reaction and Restriction Enzyme Analysis to directly detect and identify<i>Salmonella typhimurium</i>in food

Cocolin, Luca Simone; Manzano, Marisa; Cantoni, C.; Comi, Giuseppe

doi:10.1111/j.1365-2672.1998.00575.x

Cited by 39 publications

(22 citation statements)

References 18 publications

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“…Identifying which virulence factors are involved in SPI-1-independent human intestinal salmonellosis will now be an interesting and important avenue of investigation. In addition, invA was used as the molecular marker (1,3,4,5,6,13,15,16,18,19) of Salmonella diagnosis, so one should reevaluate molecular detection techniques for Salmonella that are predicated on the absolute presence of SPI-1 in humandisease-causing strains of S. enterica.…”

Section: Discussionmentioning

confidence: 99%

“…Based on the hypothesized necessity for SPI-1 in intestinal disease and its conservation among Salmonella species (14), SPI-1-encoded genes such as invA and hilA have been extensively exploited as molecular markers for the detection of enteropathogenic S. enterica (1,3,4,5,6,13,15,16,18,19). Notably, isolates of S. enterica serovars Litchfield and Senftenberg lack SPI-1 and have decreased invasion of cultured epithelial cells (9).…”

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confidence: 99%

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Salmonella enterica Serovar Senftenberg Human Clinical Isolates Lacking SPI-1

Coburn

Deng

et al. 2008

J Clin Microbiol

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Nontyphoidal Salmonella species cause gastrointestinal disease worldwide. The prevailing theory of Salmonella enteropathogenesis is that bacterial invasion of the intestinal epithelium is essential for virulence and that this requires the virulence-associated genomic region Salmonella pathogenicity island 1 (SPI-1). Recent studies of Salmonella enterica infection models have demonstrated that enterocolitis and diarrhea in mice and cows can occur independently of SPI-1. In this study, we sought to confirm whether two S. enterica serovar Senftenberg clinical isolates lacked genes essential for SPI-1 function. Two clinical strains were isolated and identified as being S. enterica serovar Senftenberg from four stool samples from a food-borne disease outbreak affecting seven individuals in Shenzhen, Guangdong Province, China, using conventional methods, pulsed-field gel electrophoresis and multilocus sequence typing. The possibility of coinfection with other potential bacteria or usual viruses was excluded. Two isolates were analyzed for the presence of invA, sipA, ssaR, sifA, and sopE2 by PCR and Southern blotting and were then assayed for the presence of SPI-1 by PCR and long-range PCR for fhlA-hilA, hilA-spaP, and spaP-invH and Southern blot analysis. A long-range PCR fragment from fhlA to mutS covering the 5 and 3 flanks of SPI-1 was also amplified from the two clinical isolates and sequenced. In addition, the two clinical isolates were assayed for enteroinvasiveness in vitro. Murine infection models were also examined. Biochemical tests and serotyping confirmed that the two clinical isolates are S. enterica serovar Senftenberg. However, they lacked genes critical for SPI-1 function but contained SPI-2 genes and were attenuated for the invasion of cultured intestinal epithelial cells. In conclusion, clinical S. enterica serovar Senftenberg strains isolated from a food-borne disease outbreak lack the invasion-associated locus SPI-1, indicating that SPI-1 is not essential for human gastroenteritis.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Salmonella enterica Serovar Senftenberg Human Clinical Isolates Lacking SPI-1

Coburn

Deng

et al. 2008

J Clin Microbiol

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“…They identified low levels of E. coli at several occupied locations, but Clostridium was only detected in a sewage outfall sample. Other potential indicator species include Enterococcus spp., Peptostreptococcus (Pre-scott et al, 1999) and Salmonella (Cocolin et al, 1998;Palmgren et al, 2000). Certain strains of these species may be specific to humans, whilst others may be found in the normal gut flora of other mammalian and avian hosts (Wang et al, 1996).…”

Section: Enteric Bacteriamentioning

confidence: 99%

PCR-based detection of non-indigenous microorganisms in ‘pristine’ environments

Baker

Tow

Cowan

2003

Journal of Microbiological Methods

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PCR-based technologies are widely employed for the detection of specific microorganisms, and may be applied to the identification of non-indigenous microorganisms in 'pristine' environments. For 'pristine' environments such as those found on the Antarctic continent, the application of these methods to the assessment of environmental contamination from human activities must be treated with caution. Issues such as the possibility of non-human dispersal of organisms, stability and survival of non-indigenous organisms in vivo, the sensitivity, reproducibility and specificity of the PCR process (and particularly primer design) and the sampling regime employed must all be considered in detail. We conclude that despite these limitations, PCR and related technologies offer enormous scope for assessment of both natural and non-indigenous microbial distributions.

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“…Furthermore, the sensitivity of the method was not increased after 8 h of enrichment on BPW (data not shown), thus allowing definition of the optimal enrichment time as 6 h. This result represents an important advance in respect to other methods which, because they prescribe longer enrichment times, slow down the detection of Salmonella spp. in food samples (3,7,8). Moreover, taking into account that, after the nonselective enrichment in BPW, about 5 to 6 h is required for the preparation of the crude extract, PCR, and electrophoresis, the detection of Salmonella spp.…”

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Twelve-Hour PCR-Based Method for Detection of Salmonella spp. in Food

Ferretti¹,

Mannazzu²,

Cocolin

et al. 2001

Appl Environ Microbiol

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A PCR-based method for the detection of Salmonella spp. in food was developed. The method, set up on typical salami from the Italian region of Marche, is sensitive and specific and shows excellent correlation with the conventional method of reference when naturally contaminated foods are analyzed. Moreover, it can be easily performed within a maximum of 12 h from food sampling, thus allowing prompt detection of Salmonella spp. in the food stocks analyzed.

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Use of Polymerase Chain Reaction and Restriction Enzyme Analysis to directly detect and identifySalmonella typhimuriumin food

Cited by 39 publications

References 18 publications

Salmonella enterica Serovar Senftenberg Human Clinical Isolates Lacking SPI-1

Salmonella enterica Serovar Senftenberg Human Clinical Isolates Lacking SPI-1

PCR-based detection of non-indigenous microorganisms in ‘pristine’ environments

Twelve-Hour PCR-Based Method for Detection of Salmonella spp. in Food

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