Multicenter validation study of two blockcycler- and one capillary-based real-time PCR methods for the detection of Salmonella in milk powder

Malorny, Burkhard; Mäde, Dietrich; Teufel, P.; Berghof-Jäger, Cornelia; Huber, Ingrid; Anderson, A. B.; Helmuth, Reiner

doi:10.1016/j.ijfoodmicro.2007.04.004

Cited by 41 publications

(20 citation statements)

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“…Moreover, validation confirms its reproducibility and specificity when used by other laboratories (Malorny et al 2003a, b). Most of the available alternative methods for detecting Salmonella in environmental samples have only been validated for the analysis of foods and feeds (Malorny et al 2009;Eijkelkamp et al 2009;Saroj et al 2008;Malorny et al 2004;Malorny et al 2007). These matrices cause less difficulty for Salmonella recovery than fecal samples, due mainly to the presence of less competitive background flora or PCR inhibitory compounds (Korsak et al 2004).…”

Section: Introductionmentioning

confidence: 87%

Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples

et al. 2009

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A comparative study of enzyme-linked fluorescent assay (ELFA)-based methods and real-time polymerase chain reaction (PCR)-based methods using three and two different sample preparation protocols, respectively, and the standard culture-based method EN ISO 6579:2002/ Amd1:2007, for the detection of Salmonella spp. in chicken feces, was performed on 20 artificially and 68 naturally contaminated chicken feces. Selectivity, relative specificity, relative accuracy, relative sensitivity, and relative detection level were determined. According to criteria established in the methods comparison study included in EN ISO 16140:2003 for validation of alternative microbiological methods, the ELFA-based methods (V1 and V2) as well as a real-time PCR method (PCR2) were comparable to the reference method for the detection of Salmonella in chicken feces. They provided results in 48 h and presented a high sensitivity (97% for all of them). The three methods showed a relative specificity of 94%, V1 being the method which presented the highest relative accuracy (96%). While detection level for V2 and reference method was between 3 and 13 CFU/25 g, PCR2 method was able to detect down to 3 CFU/25 g. In conclusion, both the real-time PCR and the ELFA-based assays can be used as rapid and user-friendly screening methods for detection of Salmonella spp. in chicken feces.

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Section: Introductionmentioning

confidence: 87%

Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples

et al. 2009

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“…Meanwhile, more and more laboratories introduced the ttr real-time PCR assay (Malorny et al 2007d). The assay was extensively tested in respect to its detection limit, inclusivity, exclusivity, specificity, sensitivity, and robustness.…”

Section: Real-time Pcr Assays For the Detection Of Salmonellamentioning

confidence: 99%

“…The other assay uses primers and a TaqMan probe located in the ttr locus according to Malorny et al (2004b). An inter-laboratory study has shown the validity of the methods (Malorny et al 2007d). …”

Section: Validation and Standardization Of Pcr For The Detection Of Smentioning

confidence: 99%

“…Content and cycle conditions of typical ttr real-time PCR reaction according toMalorny et al (2007d) Available from Applied Biosystems, Foster City, CA, USA. Other mastermixes such as Quantitect Multiplex PCR Kit (Qiagen, Hilden, Germany) or Brilliant Multiplex QPCR Master Mix (Stratagene, USA) also possible b Available from the National Salmonella Reference Laboratory at Federal Institute for Risk Assessment, Berlin, Germany…”

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confidence: 99%

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Polymerase Chain Reaction for the Rapid Detection and Serovar Identification of Salmonella in Food and Feeding Stuff

et al. 2008

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The polymerase chain reaction (PCR) is one of the most important rapid methods for the sensitive and specific detection of pathogenic and spoilage microorganisms. The method is increasingly applied in surveillance and monitoring programs to detect pathogens, especially for ensuring the safety and quality of food. The food-borne pathogen Salmonella together with Campylobacter is the most predominant bacterial pathogen in Europe, causing approximately 160,000 confirmed human cases per year. During the last two decades, the importance of Salmonella for food safety triggered the development of dozens of PCR-based detection methods. They promise significant advantages compared to the traditional culture-based methods with respect to speed, easiness, reliability, sensitivity, cost effectiveness, and automation. However, many of them are not applicable because of lacking validation procedures. Meanwhile, PCR has internationally been standardized and guidelines as well as standards for the validation of alternative methods, such as PCR, were established. This review will give an overview of the historical development of PCR-based methods for the detection of Salmonella including validation aspects and summarizes the state-of-the-art for the detection of Salmonella in food and feeding stuff by realtime PCR. Furthermore, current multiplex PCR-based serovar-specific identification methods will be reviewed.

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“…Thereby favoring SE targets instead of the DNA IAC. Nevertheless, it is important to emphasize that performance of the IAC in the presence of low DNA copy numbers or in the observed absence of SE was robust and reliable for each food sample analyzed, an imperative finding for any Salmonella detection qPCR assay (Malorny et al, 2007b).…”

Section: Discussionmentioning

confidence: 99%

Multiplex TaqMan Real-Time PCR (qPCR) Assay Targeting prot6E and invA Genes for Fast and Accurate Detection of Salmonella Enteritidis

González-Escalona¹,

Zhang²,

Brown³

2012

Salmonella - A Diversified Superbug

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Multicenter validation study of two blockcycler- and one capillary-based real-time PCR methods for the detection of Salmonella in milk powder

Cited by 41 publications

References 3 publications

Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples

Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples

Polymerase Chain Reaction for the Rapid Detection and Serovar Identification of Salmonella in Food and Feeding Stuff

Multiplex TaqMan Real-Time PCR (qPCR) Assay Targeting prot6E and invA Genes for Fast and Accurate Detection of Salmonella Enteritidis

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