Identification by a multiplex PCR‐based assay of<i>Salmonella</i>Typhimurium and<i>Salmonella</i>Enteritidis strains from environmental swabs of poultry houses

Soumet, Christophe; Ermel, Gwennola; Rose, Valérie; Rose, Nicolas; Drouin, P.; Salvat, Gilles; Colin, P.

doi:10.1046/j.1365-2672.1999.00559.x

Cited by 95 publications

(81 citation statements)

References 22 publications

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“…PCR with preincubation in an enrichment broth has been performed for human (5,13,14,28), animal (6,23,24), fecal, and food (1,2,4,9,11,19) samples. This has been found to be a useful and more rapid method because it increases the number of viable Salmonella in the sample and, therefore, increases the sensitivity of the assay (5,9,11,22). PCR methods and DNA extraction procedures for Salmonella have been described for human, animal, and food samples (5,6,21,23,24).…”

mentioning

confidence: 99%

Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Çarlı¹,

Unal²,

Caner³

et al. 2001

J Clin Microbiol

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This report describes a rapid detection procedure for salmonellae from chicken feces by the combination of tetrathionate primary enrichment (preenrichment [PE])-bacterial lysis-capillary PCR and capillary gel electrophoresis. Pure Salmonella enterica serovar Enteritidis 64K was reisolated and detected by capillary PCR after buffered peptone water and nutrient broth, tetrathionate broth base Hajna (TTBH), and tetrathionate broth (TTB) preenrichments. When the same culture was mixed with intestinal homogenate, bacteriological reisolation and capillary PCR detection was achieved only by TTBH and TTB preenrichments. Capillary gel electrophoresis revealed that a Salmonella genus-specific 281-bp PCR product was detected when Salmonella strains but not non-Salmonella strains were tested. The detection limit of capillary PCR with whole-cell DNA extracted from pure Salmonella enterica serovars Enteritidis 64K, Typhimurium LT2-CIP60-62, and Gallinarum 64K was 3, 3, and 9 CFU ml ؊1 , respectively. The detection limit of capillary PCR from whole-cell DNA extracted from intestinal homogenate artificially contaminated with the same three strains was 3, 3, and 7 CFU ml ؊1 , respectively. We compared the results of the capillary PCR and bacteriological examination from the natural samples. Thirty-five of 53 naturally contaminated samples produced a specific PCR product. In 9 of the 35 PCR-positive samples, Salmonella could not be detected bacteriologically either by PE or a primary and delayed secondary enrichment (DSE) combination. In the 18 PCR-negative samples, 4 samples were found to harbor Salmonella by both PE and DSE and 14 samples were positive after DSE. Fifty-three additional intestinal homogenate samples, which were negative by their PE and DSE in bacteriological examination, were found to be also negative by their PCRs. The total time required to detect Salmonella with the capillary PCR method we used was approximately 20 h. If samples are from clinically diseased birds, the total time for PCR and detection is reduced to 2 h since the 18-h PE is not required. These results indicate that TTB enrichment, bacterial lysis, and genus-specific capillary PCR combined with capillary gel electrophoresis constitute a sensitive and selective procedure which has the potential to rapidly identify Salmonella-infected flocks.Salmonellae are among the major bacterial pathogens of poultry in Turkey and in the world (3,8). Prevention of Salmonella infection is important for poultry health and for the food industry, and prevention can be achieved only by good monitoring and screening programs (10,17,27). Salmonella detection by bacteriological methods usually requires 5 to 11 days (10, 27), and samples with low numbers of Salmonella cells, usually seen in subclinically infected chickens, may give false-negative results (7). Efforts have been made to reduce the time required and to increase the sensitivity of methods to detect Salmonella serovars in poultry samples (16,25). PCR with preincubation in an enrichment broth has been perform...

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confidence: 99%

Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Çarlı¹,

Unal²,

Caner³

et al. 2001

J Clin Microbiol

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“…It must be considered however, that the majority of the articles in the scientific literature deal with mPCR methods developed to identify and or characterize Salmonella serotypes from pure cultures, or in controlled artificial inoculation experiments, with only a minority of studies providing results from in situ detection of pathogens in foods or environmental samples. Soumet et al (1999) developed a multiplex PCR assay for the simultaneous identification of Salmonella species, S. Enteritidis and S. Typhimurium from environmental swabs of poultry houses. Similarly, O'Regan et al (2008) developed a real-time multiplex PCR assay for the detection of multiple Salmonella serotypes in chicken samples.…”

Section: Multiplex Pcrmentioning

confidence: 99%

Salmonella Detection Methods for Food and Food Ingredients

Odumeru¹,

León-Velarde²

2012

Salmonella - A Dangerous Foodborne Pathogen

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“…Traditional methods for Salmonella detection in foods are based on culture using selective media (enrichment) and further identification of suspect colonies by biochemical and serological testing (Soumet et al 1999). These methods are labour-intensive and time-consuming, usually taking about 4-6 days to achieve confirmed positive test results.…”

Section: Introductionmentioning

confidence: 99%

Nucleic acid sequence based amplification for the rapid and sensitive detection of Salmonella enterica from foods

D’Souza

Jaykus

2003

J Appl Microbiol

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Aims: The purpose of this study was to apply nucleic acid sequence-based amplification (NASBA) for the detection of Salmonella enterica serovar Enteritidis (S. Enteritidis) in representative foods. Methods and Results: A previously reported primer and probe set based on mRNA sequences of the dnaK gene of Salmonella were used in this study. To test for possible food matrix inhibition and assay detection limits, 25-g samples of representative food commodities (fresh meats, poultry, fish, ready-to-eat salads and bakery products) were pre-enriched with and without S. Enteritidis inoculation. The NucliSens Ò Basic Kit, supplemented with enzymes from various other commercial sources, was used for RNA isolation, NASBA amplification and electrochemiluminescent (ECL) detection. The end point detection limit of the NASBA-ECL assay was equivalent to 10 1 CFU of S. Enteritidis per amplification reaction. When the assay was tested on noncontaminated foods, none of the food matrices produced false-positive results. Some of the food matrices inhibited the NASBA-ECL reaction unless the associated RNA was diluted 10-fold prior to amplification. Conclusions: For all food items tested, positive ECL signals were achieved after 18 h of pre-enrichment and subsequent NASBA at initial inoculum levels of 10 2 and 10 1 CFU per 25 g food sample.Significance and Impact of the Study: This rapid, semi-automated detection method has potential for use in the food, agricultural and public health sectors.

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Identification by a multiplex PCR‐based assay ofSalmonellaTyphimurium andSalmonellaEnteritidis strains from environmental swabs of poultry houses

Cited by 95 publications

References 22 publications

Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Salmonella Detection Methods for Food and Food Ingredients

Nucleic acid sequence based amplification for the rapid and sensitive detection of Salmonella enterica from foods

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