Direct detection of Salmonella spp. in estuaries by using a DNA probe

Knight, Ivor T.; Shults, S; Kaspar, Charles W.; Colwell, R. R.

doi:10.1128/aem.56.4.1059-1066.1990

Cited by 65 publications

(22 citation statements)

References 31 publications

(39 reference statements)

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“…In this study, we found that real-time PCR detection sensitivity from natural samples was higher than bacteriology both in intestinal samples and in total, when the samples taken from each¯ock were considered individually (Table 1). Possible reasons for this can be as follows: salmonellae found in natural samples may show atypical biochemical pro®les and may not be detected in bacteriology (Bennet et al 1998), and Salmonella cells may be present in a viable but non-culturable status (Knight et al 1990). The number of Salmonella-positive¯ocks was 13 (27á7%) both by PCR and culture method.…”

Section: Resultsmentioning

confidence: 99%

Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry

Eyigör

Çarlı

Unal³

2002

Lett Appl Microbiol

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Aims: The present study describes the implementation of real‐time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry. Methods and Results: Real‐time PCR with Salmonella invA‐specific primers and a standard bacteriological method was applied to detect Salmonella in tetrathionate enrichment cultures of 492 intestinal homogenates and 27 drag swabs from 47 poultry flocks. The number of positive individual samples by real‐time PCR and culture method was 65 (12·5%) and 35 (6·8%), respectively. The number of Salmonella‐positive flocks was 13 (27·7%) by both methods. PCR detection required 25 min for up to 32 samples. Melting curve analysis revealed the Tm for Salmonella‐specific PCR product as 87 ± 1°C. Conclusions: Implementation of real‐time PCR to tetrathionate broth enrichment step reduces the Salmonella detection time to 18 h and 25 min. Isolation of Salmonella should be carried out with PCR to determine the serovar. Significance and Impact of the Study: Real‐time PCR is a powerful tool in rapid and accurate Salmonella monitoring in poultry companies, together with standard bacteriology.

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Section: Resultsmentioning

confidence: 99%

Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry

Eyigör

Çarlı

Unal³

2002

Lett Appl Microbiol

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“…In this study, the number of positive individual samples by real-time PCR was almost double that detected by the culture method. A possible explanation of these results is that salmonellae found in natural samples may show atypical biochemical profiles and may not be detected using bacteriological methods, and that Salmonella cells may be present in a viable but nonculturable state (Knight et al 1990;Bennett et al 1998;Eyigor et al 2002). In recent years, several studies have been published in which the use of a conventional PCR assay proved more sensitive than the culture method for detecting Salmonella in poultry, meat, and poultry-related products (Bennett et al 1998;Whyte et al 2002;Oliveira et al 2002;Fratamico 2003).…”

Section: Discussionmentioning

confidence: 99%

Adding a selective enrichment step to the iQ-CheckTM real-time PCR improves the detection of Salmonella in naturally contaminated retail turkey meat products

Fakhr¹,

McEvoy²,

Sherwood³

2006

Lett Appl Microbiol

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Aims: The aim of this study was to compare the real‐time iQ‐CheckTMSalmonella kit (Bio‐Rad) with the immunocapture assay RapidCheck®Salmonella method, and a conventional culture method (FSIS, USDA) in detecting Salmonella in naturally contaminated turkey meat products. This study was also designed to determine if a selective enrichment step might improve the real‐time detection of Salmonella. Methods and Results: Using the culture method, Salmonella was recovered from 49 out of 99 retail turkey meat samples collected. RapidCheck® failed to detect 11 Salmonella samples that were positive by the culture method. The iQ‐CheckTM real‐time PCR also failed to detect three samples that were positive by the culture method. However, when carried out after a selective enrichment step, the iQ‐CheckTM real‐time PCR detected all 49 Salmonella samples recovered by the culture method. The iQ‐CheckTM real‐time PCR detected the presence of Salmonella in some samples that were not recovered by the culture method. Conclusions: Adding a selective enrichment step to the iQ‐CheckTM real‐time PCR improves the detection of Salmonella in naturally contaminated turkey meat samples. Significance and Impact of the Study: The iQ‐CheckTMSalmonella real‐time PCR can be used as a rapid method to monitor Salmonella in turkey meat, together with conventional culture methodology.

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“…Although the DNA-DNA hybridization method using Salmonella-specific DNA probes offers one of the rapid methods for Salmonella detection (Fitts et ul. 1983;Tsen et al 1989Tsen et al , 1991Knight et al 1990;Cano et al 1992), this method requires enrichment steps for target cells and thus, about 48-72 h are needed to complete the detection (Emswiler-Rose et al 1987;Tsen et al 1991). The polymerase chain reaction (PCR) technique offers another possibility for rapid detection of Salmonella.…”

Section: Introductionmentioning

confidence: 99%

“…Although 23s rRNA targeted DNA probes have been developed (Ronner and Stackebrandt 1994) and are commercially available (Knight et al 1990;Wilson et al 1990b), to our knowledge, 16s RNA gene-based PCR primers have not been developed for the specific detection of Salmonella in foods. Recently, a rapid method for the detection of Salmonella strains using a combination of PCR and reverse dot-blot hybridization has been reported (Iida et al 1993).…”

Section: Introductionmentioning

confidence: 99%

Use of two 16S DNA targeted oligonucleotides as PCR primers for the specific detection of Salmonella in foods

Lin

Tsen

1996

Journal of Applied Bacteriology

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A 16S DNA targeted polymerase chain reaction (PCR) method specific for the detection of Salmonella isolates with various serotypes was developed. The primers used for such a PCR method were 16SF1 and 16SIII. 16SF1 is the reverse and complementary strand of 16SI which has been shown to be able to hybridize with Salmonella and Citrobacter spp. 16III on the other hand, is able to hybridize with Klebsiella and Serratia spp. in addition to Salmonella. Although 16SF1 and 16SIII were not specific to Salmonella only, when they were used as PCR primers, only the Salmonella isolates could be specifically detected. The interference from Citrobacter, Klebsiella and Serratia spp. could be prevented. None of the other non-Salmonella isolates including strains of the family of Enterobacteriaceae closely related to Salmonella would generate the false-positive reaction. When this PCR system was used for the detection of Salmonella cells artificially contaminated in food samples, results obtained were satisfactory. A detection limit of N x 10(0) cells per assay could be obtained.

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Direct detection of Salmonella spp. in estuaries by using a DNA probe

Cited by 65 publications

References 31 publications

Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry

Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry

Adding a selective enrichment step to the iQ-CheckTM real-time PCR improves the detection of Salmonella in naturally contaminated retail turkey meat products

Use of two 16S DNA targeted oligonucleotides as PCR primers for the specific detection of Salmonella in foods

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