Multiplex real-time PCR for the detection and quantification of DNA from duck, goose, chicken, turkey and pork

Köppel, René; Daniels, Michael; Felderer, Nora; Brünen-Nieweler, Claudia

doi:10.1007/s00217-013-1973-2

Cited by 38 publications

(29 citation statements)

References 11 publications

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“…Contradicting the vendors' declarations, significant amounts of pork were detected using both methods. It has been reported previously that the usage of relatively cheap meat such as pork is common in the manufacture of pt (11); however, with the samples tested here, the relative amount of pork is even higher than the amount of meat the samples are declared to be manufactured from: duck, chicken, and pheasant (Sample 1) and goose (Sample 2), respectively.…”

Section: Resultsmentioning

confidence: 79%

“…For the food panel, most values of multiplex MP PCR compared with multiplex HP PCR from Microsynth are within the acceptable range according to FDA guidelines (www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm368107.pdf) as well as within the range of previously published data (11) (Table 2). …”

Section: Resultsmentioning

confidence: 90%

“…The sequences of primers and probes for food panel and methicillin-resistant Staphylococcus aureus (MRSA) panel detection are based on the literature (11,12). All oligonucleotides were purchased from biomers.net (Ulm, Germany) except for primers and probes for the HP PCR-based food detection panel, which were sourced from a commercial kit (Pentaplex Real-Time PCR AllPat, Microsynth, Switzerland).…”

Section: Methodsmentioning

confidence: 99%

“…Label-free MPs based on the respective HP sequences were easily designed according to recently described guidelines (11,12). Briefly, the 5-terminal fluorophore of each HP was replaced by a 17-nucleotide generic sequence (18 nucleotides in the case of MP PVL ) that is the reverse-complement to the MP hybridization sequence of one of the URs.…”

Section: Methodsmentioning

confidence: 99%

“…The set was then validated using two different multiplex real-time MP PCR assays (Figure 1). The first assay is based on a pentaplex HP PCR food panel that discriminates DNA sequences from different animal species potentially present in pt, a speciality food product originally made from goose or duck (11). Instead of the five dual-labeled HPs of the original assay, five label-free MPs together with this UR set were used.…”

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confidence: 99%

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Simplified Development of Multiplex Real-Time PCR Through Master Mix Augmented by Universal Fluorogenic Reporters

et al. 2016

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Mediator probe (MP) PCR is a real-time PCR approach that uses standardized universal fluorogenic reporter oligonucleotides (UR) in conjunction with label-free sequence-specific probes. To enable multiplex real-time MP PCR, we designed a set of five optimized URs with different fluorescent labels. Performance of the optimized URs was verified in multiplex real-time MP PCR for the detection of a pentaplex food panel and a quadruplex methicillin-resistant Staphylococcus aureus (MRSA) panel. Results were comparable to corresponding multiplex hydrolysis probe (HP) PCR, also designated as TaqMan PCR. Analyses of MRSA DNA standards and DNA extracted from patient swab samples showed improved lower limits of detection (LoDs) by a factor of 2-5 when using quadruplex real-time MP PCR instead of HP PCR. The novel set of standardized URs we present here simplifies development of multiplex real-time PCR assays by requiring only the design of label-free probes. In the future, real-time PCR master mixes could be augmented with up to five standardized fluorogenic URs, each emitting light at a different wavelength.

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Section: Resultsmentioning

confidence: 79%

Section: Resultsmentioning

confidence: 90%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Simplified Development of Multiplex Real-Time PCR Through Master Mix Augmented by Universal Fluorogenic Reporters

et al. 2016

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Meat authenticity

Zhao

2016

Emerging Technologies in Meat Processing

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Improved triplex real‐time PCR with endogenous control for synchronous identification of DNA from chicken, duck, and goose meat

Liu¹,

Luo²,

Xu³

et al. 2021

Food Science & Nutrition

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The authentication and labeling of meat products, concerning origins and species, are key to fair trade and to protect consumer interests in the market. We developed an improved triplex real‐time PCR approach to simultaneously identify chicken, duck, and goose DNA in meat, including an endogenous control to avoid false negatives. Our method specifically detected DNA from chicken, duck, and goose, and showed no cross‐reaction with DNA extracted from other meat types. The detection limits of chicken, duck, and goose DNA were 0.001–0.00025 ng, 0.0025–0.0001 ng, and 0.001–0.00001 ng, respectively, and we were able to simultaneously identify DNA from two distinct origins using as little as 0.1% of total meat weight. Our newly generated triplex real‐time PCR method with endogenous control exhibited high specificity, sensitivity, and efficiency for simultaneous identification of DNA from chicken, duck, and goose in meat.

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Multiplex real-time PCR for the detection and quantification of DNA from duck, goose, chicken, turkey and pork

Cited by 38 publications

References 11 publications

Simplified Development of Multiplex Real-Time PCR Through Master Mix Augmented by Universal Fluorogenic Reporters

Simplified Development of Multiplex Real-Time PCR Through Master Mix Augmented by Universal Fluorogenic Reporters

Meat authenticity

Improved triplex real‐time PCR with endogenous control for synchronous identification of DNA from chicken, duck, and goose meat

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