Detection of pathogenic and spoilage micro-organisms in food with the polymerase chain reaction

Scheu, P.; Berghof, K.; Ståhl, Ulf

doi:10.1006/fmic.1997.0134

Cited by 170 publications

(100 citation statements)

References 125 publications

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“…In addition, the use of enrichment promotes germination and outgrowth of clostridial spores and, consequently, improves the yield of extracted DNA. PCR has demonstrated a wide potential for specific recognition of bacteria without their prior enrichment (Scheu et al 1998). However, PCR detects the presence of target DNA in the sample, rather than the presence of viable and/or culturable microorganisms.…”

Section: Discussionmentioning

confidence: 99%

Abattoir sources of psychrophilic clostridia causing blown pack spoilage of vacuum-packed chilled meats determined by culture-based and molecular detection procedures

Boerema

Broda

Bell

2003

Letters in Applied Microbiology

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Aims: To identify the abattoir source(s) of psychrophilic clostridia causing 'blown pack' spoilage of vacuum-packed chilled meats. Methods and Results: Molecular procedures were used to detect the presence of specific 16S rRNA gene fragments of blown pack-causing clostridia in samples collected from a commercial abattoir and its environs. Blown pack-causing clostridia were consistently detected in hide, soil and faecal samples, as well as in samples collected at slaughter plant locations associated with handling of animals and animal carcasses prior to pelt removal. Conclusions: The data indicate that pelts per se or soil particles/faecal material attached thereto are the most probable primary reservoir of blown pack clostridia in the abattoir. Significance and Impact of the Study: The paper provides information critical for controlling blown pack spoilage in commercial meat-processing plants.

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Section: Discussionmentioning

confidence: 99%

Abattoir sources of psychrophilic clostridia causing blown pack spoilage of vacuum-packed chilled meats determined by culture-based and molecular detection procedures

Boerema

Broda

Bell

2003

Letters in Applied Microbiology

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“…Hill, 1996;Lantz et al, 2000;Olsen et al, 1995;Scheu et al, 1998). PCRbased methods are predicted to be established as routine reference methods within the next ten years , however further developments are needed for an effective implementation of this technique in food microbiology.…”

Section: Pcr-based Methodsmentioning

confidence: 99%

“…Although several methodological approaches have been described based on conventional PCR (Rijpens and Hermann, 2002;Scheu et al, 1998), the most promising alternative is the application of adequate RTi-PCR assays.…”

Section: Pcr-based Methodsmentioning

confidence: 99%

“…viable but non-culturable VBNC forms). A number of alternative, rapid and sensitive methods for the detection, identification and quantification of foodborne pathogens have been developed to overcome these drawbacks, including bioluminescence-based, immunological and molecular methods (Fung, 2002;Scheu et al, 1998). Among the latter category, PCR has become the most popular microbiological diagnostic method, and recently, the introduction of a development of this technique, RTi-PCR, has produced a second revolution in the molecular diagnostic methodology in microbiology as stated by the increasing number of scientific publications and novel commercial kits available.…”

Section: ____________________________________________________________mentioning

confidence: 99%

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Molecular methodology in Food Microbiology diagnostics: trends and current challenges

Rodrı́guez-Lázaro

Hernández

2006

13th World Congress of Food Science &Amp; Technology

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Foodborne diseases are among the most serious public health concerns worldwide. Consequently, the application of microbiological controls within the quality assessment programs in the food industry is a premise to minimize the risk of infection for the consumer. Classical microbiological methods involve, in general, the use of appropriate pre-enrichment and enrichment, isolation on selective media, and subsequent confirmation using morphological, biochemical and/or serological tests. They are laborious, time consuming and not always reliable. A number of alternative, rapid and sensitive molecular methods for the detection, identification and quantification of foodborne pathogens have been developed to overcome these drawbacks. However, until now, they have shown low practical implementation for monitoring and microbiological control. Amplification techniques have a number of advantages including improved speed, excellent detection limit, specificity, sensitivity and potential for quantification as well as easy automation. Major drawbacks are the cost of the analysis and the qualified personnel needed to carry out the experiments, plus the lack of standardisation of protocols approved by normalisation bodies. Moreover, the cost of reagents is decreasing and more qualified people are available to perform this kind of analysis. Efforts on standardisation and normalisation of these techniques are a key priority.The molecular methods, in conclusion, are a promising alternative that can substitute or complement the current reference methods in food microbiology diagnostics. More suitable and reliable results can be achieved in terms of speed and precision, and can detect and enumerate specifically viable microorganisms.

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“…Furthermore, the absence of post-PCR analytical steps reduces cross-contamination risks and allows high throughput and automation (Klein, 2002;Mackay, 2004). However, many components of food products, culture media and nucleic acids extraction reagents can act as PCR inhibitors Rossen et al, 1992; Wilson, 1997), and their presence in the reaction may cause a dramatic decrease in sensitivity or even block amplification reaction, consequently generating false negative results or an underestimation of the bacterial load (Hill, 1996;Scheu et al, 1998;Rådström et al, 2003). Therefore, the assessment of the PCR efficiency in every assay is a critical issue for the implementation of PCR as a routinely analytical tool in food microbiology diagnostics.…”

Section: Introductionmentioning

confidence: 99%

An Optimised Quantitative Real-Time PCR Assay for Listeria Monocytogenes Including and Internal Amplification Control

Rodrı́guez-Lázaro

Pla

Scortti

et al. 2006

13th World Congress of Food Science &Amp; Technology

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The assessment of results obtained by PCR is a critical issue for its implementation as a routine tool in food microbiology diagnostics. As an analytical technique, PCR is exposed to inhibitors that can produce false negative results or underestimation in quantitative analysis. In this study, we illustrate the design, development and optimization of a duplex real-time PCR (RTi-PCR) assay for the quantitative detection of Listeria monocytogenes based on the co-amplification of a L. monocytogenes-specific gene (hly) sequence (Rodríguez-Lázaro et al., 2004a) and an internal amplification control (IAC) to evaluate PCR performance. A PCR IAC is a non-target DNA that is co-amplified with the same set of primers that target sequence. The assay was 100% specific as it allowed unambiguously detection of 49 L. monocytogenes isolates of different serotypes and sources and 104 strains of other species were negative. The detection and quantification limits were 8 and 30 cfu respectively, and PCR linearity and efficiency were R 2 =0.997 and 0.80 respectively. Interestingly, the assay was capable of detecting underestimation of L. monocytogenes in reactions containing inhibitory culture media (e.g. Fraser and Half Fraser media) and foodstuffs (e.g. raw salmon or raw pork meat). Specific L. monocytogenes detection could also be performed using agargrown colonies and by conventional PCR. In conclusion, we present a simple and rapid duplex RTi-PCR assay for the quantitative detection of L. monocytogenes, which can also assess potential underestimations or false negative results.

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Detection of pathogenic and spoilage micro-organisms in food with the polymerase chain reaction

Cited by 170 publications

References 125 publications

Abattoir sources of psychrophilic clostridia causing blown pack spoilage of vacuum-packed chilled meats determined by culture-based and molecular detection procedures

Abattoir sources of psychrophilic clostridia causing blown pack spoilage of vacuum-packed chilled meats determined by culture-based and molecular detection procedures

Molecular methodology in Food Microbiology diagnostics: trends and current challenges

An Optimised Quantitative Real-Time PCR Assay for Listeria Monocytogenes Including and Internal Amplification Control

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