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Margit Cichnamarkl - One of the best experts on this subject based on the ideXlab platform.

  • real time pcr assay for the detection and quantification of roe deer to detect Food Adulteration interlaboratory validation involving laboratories in austria germany and switzerland
    Foods, 2021
    Co-Authors: Barbara Druml, Margit Cichnamarkl, Steffen Uhlig, Kirsten Simon, Kirstin Frost, Karina Hettwer, Rupert Hochegger
    Abstract:

    Game meat products are particularly prone to be adulterated by replacing game meat with cheaper meat species. Recently, we have presented a real-time polymerase chain reaction (PCR) assay for the identification and quantification of roe deer in Food. Quantification of the roe deer content in % (w/w) was achieved relatively by subjecting the DNA isolates to a reference real-time PCR assay in addition to the real-time PCR assay for roe deer. Aiming at harmonizing analytical methods for Food authentication across EU Member States, the real-time PCR assay for roe deer has been tested in an interlaboratory ring trial including 14 laboratories from Austria, Germany, and Switzerland. Participating laboratories obtained aliquots of DNA isolates from a meat mixture containing 24.8% (w/w) roe deer in pork, roe deer meat, and 12 meat samples whose roe deer content was not disclosed. Performance characteristics included amplification efficiency, level of detection (LOD95%), repeatability, reproducibility, and accuracy of quantitative results. With a relative reproducibility standard deviation ranging from 13.35 to 25.08% (after outlier removal) and recoveries ranging from 84.4 to 114.3%, the real-time PCR assay was found to be applicable for the detection and quantification of roe deer in raw meat samples to detect Food Adulteration.

  • red deer cervus elaphus specific real time pcr assay for the detection of Food Adulteration
    Food Control, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    Abstract We present a red deer-specific real-time PCR assay which, combined with a reference real-time PCR assay published previously, allows the quantification of the red deer content in Food products. Thus, it can be applied to detect Food Adulteration. The primer/probe system of the red deer-specific real-time PCR assay amplifies a 87 bp long fragment of the protein kinase C iota gene. To eliminate cross-reactivity with closely related species, the forward primer was designed to contain one deliberate base mismatch adjacent to one red deer-specific base. The red deer-specific real-time PCR assay did not show cross-reactivity with 23 animal and 50 plant species tested. LOD and LOQ, determined by analyzing a serially diluted DNA extract containing 1% (w/w) red deer DNA in pig DNA, were 0.05% and 0.4%, respectively. The accuracy was validated by analyzing DNA mixtures, meat extract mixtures, meat mixtures and model game sausages with known red deer content. The highest accuracy was obtained when the calibration mixture was similar to the analyzed sample in both the composition and concentration of the animal species of interest. High recoveries were not only obtained for raw samples but also after subjection to thermal treatment, including brewing (15 min at 75–78 °C), boiling (90 min at 100 °C) and microwave treatment (15 s, 40 s or 2 min at 650 W). The red deer-specific real-time PCR assay was found to be robust with respect to small deviations in the reaction volume or the annealing temperature and the use of another real-time PCR instrument.

  • development and validation of a fallow deer dama dama specific taqman real time pcr assay for the detection of Food Adulteration
    Food Chemistry, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    The aim of the present study was to develop a real-time PCR assay for the identification and quantification of fallow deer (Dama dama) in Food to detect Food Adulteration. Despite high sequence homology among different deer species, a fallow deer-specific primer/probe system targeting a fragment of the nuclear MC1-R gene was designed. This primer/probe system did not amplify DNA from 19 other animals and 50 edible plant species. Moderate cross-reactivity was observed for sika deer, red deer, roe deer, reindeer and wild boar. The LOD and LOQ of the real-time PCR assay were 0.1% and 0.4%, respectively. To validate the assay, DNA mixtures, meat extract mixtures, meat mixtures and model game sausages were analyzed. Satisfactory quantitative results were obtained when the calibration mixture was similar to the analyzed sample in both the composition and concentration of the animal species of interest.

  • a novel reference real time pcr assay for the relative quantification of game meat species in raw and heat processed Food
    Food Control, 2016
    Co-Authors: Barbara Druml, Rupert Hochegger, Maria Kaltenbrunner, Margit Cichnamarkl
    Abstract:

    Abstract In order to increase their profit, Food producers may be tempted to replace expensive meat species by cheaper ones. Due to their selectivity and sensitivity, PCR based methods are frequently applied to identify and quantify meat species to detect Food Adulteration. Quantification of meat species in highly processed Food products is, however, a difficult task. When we used a previously published reference system to relatively quantify the deer content in heat treated Foodstuffs, we obtained recoveries that were substantially >100%. In the present study we aimed to improve the applicability of the reference system, targeting a 97 bp fragment of the myostatin gene, to heat treated products by reducing the length of the target sequence. The novel reference system, targeting a 70 bp fragment of the myostatin gene, was found to amplify the target region in 27 mammals and poultry species. Meat mixtures as well as raw and heat treated model sausages were analysed to demonstrate the applicability of the novel reference system for relatively quantifying the game meat content in processed Food products. With the novel reference system, the bias introduced by heat treatment was largely eliminated.

  • authenticity control of game meat products a single method to detect and quantify Adulteration of fallow deer dama dama red deer cervus elaphus and sika deer cervus nippon by real time pcr
    Food Chemistry, 2015
    Co-Authors: Barbara Druml, Stephanie Grandits, Walter Mayer, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    This contribution presents a single real-time PCR assay allowing the determination of the deer content (the sum of fallow deer (Dama dama), red deer (Cervus elaphus) and sika deer (Cervus nippon)) in meat products to detect Food Adulteration. The PCR assay does not show cross-reactivity with 20 animal species and 43 botanical species potentially contained in game meat products. The limit of quantification is 0.5% for fallow deer and red deer and 0.1% for sika deer. The deer content in meat products is determined by relating the concentration obtained with the deer PCR assay to that obtained with a reference system which amplifies mammals and poultry DNA. The analysis of binary meat mixtures with pork, a meat mixture containing equal amounts of fallow deer, red deer and sika deer in pork and a model game sausage showed that the quantification approach is very accurate (systematic error generally <25%).

Rupert Hochegger - One of the best experts on this subject based on the ideXlab platform.

  • real time pcr assay for the detection and quantification of roe deer to detect Food Adulteration interlaboratory validation involving laboratories in austria germany and switzerland
    Foods, 2021
    Co-Authors: Barbara Druml, Margit Cichnamarkl, Steffen Uhlig, Kirsten Simon, Kirstin Frost, Karina Hettwer, Rupert Hochegger
    Abstract:

    Game meat products are particularly prone to be adulterated by replacing game meat with cheaper meat species. Recently, we have presented a real-time polymerase chain reaction (PCR) assay for the identification and quantification of roe deer in Food. Quantification of the roe deer content in % (w/w) was achieved relatively by subjecting the DNA isolates to a reference real-time PCR assay in addition to the real-time PCR assay for roe deer. Aiming at harmonizing analytical methods for Food authentication across EU Member States, the real-time PCR assay for roe deer has been tested in an interlaboratory ring trial including 14 laboratories from Austria, Germany, and Switzerland. Participating laboratories obtained aliquots of DNA isolates from a meat mixture containing 24.8% (w/w) roe deer in pork, roe deer meat, and 12 meat samples whose roe deer content was not disclosed. Performance characteristics included amplification efficiency, level of detection (LOD95%), repeatability, reproducibility, and accuracy of quantitative results. With a relative reproducibility standard deviation ranging from 13.35 to 25.08% (after outlier removal) and recoveries ranging from 84.4 to 114.3%, the real-time PCR assay was found to be applicable for the detection and quantification of roe deer in raw meat samples to detect Food Adulteration.

  • Tetraplex real-time PCR assay for the simultaneous identification and quantification of roe deer, red deer, fallow deer and sika deer for deer meat authentication.
    Food Chemistry, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichna-markl
    Abstract:

    Abstract Analytical methods are needed for the identification and quantification of meat species to detect Food Adulteration. Since game meat is more expensive than meat from domesticated animal species, it is a potential target for Adulteration. We present a tetraplex real-time PCR assay that allows the simultaneous determination of the content of roe deer, red deer, fallow deer and sika deer. The tetraplex assay showed only moderate cross-reactivity with closely related species. After optimization the tetraplex assay had a limit of detection of 0.1% (w/w) and a limit of quantification of 0.5% (w/w) for each of the four deer species. The tetraplex assay was found to be robust, slight modifications of the experimental setup did not lower its performance. Recoveries obtained by analyzing DNA mixtures and DNA isolates from model game sausages were similar to those obtained with the singleplex assays.

  • red deer cervus elaphus specific real time pcr assay for the detection of Food Adulteration
    Food Control, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    Abstract We present a red deer-specific real-time PCR assay which, combined with a reference real-time PCR assay published previously, allows the quantification of the red deer content in Food products. Thus, it can be applied to detect Food Adulteration. The primer/probe system of the red deer-specific real-time PCR assay amplifies a 87 bp long fragment of the protein kinase C iota gene. To eliminate cross-reactivity with closely related species, the forward primer was designed to contain one deliberate base mismatch adjacent to one red deer-specific base. The red deer-specific real-time PCR assay did not show cross-reactivity with 23 animal and 50 plant species tested. LOD and LOQ, determined by analyzing a serially diluted DNA extract containing 1% (w/w) red deer DNA in pig DNA, were 0.05% and 0.4%, respectively. The accuracy was validated by analyzing DNA mixtures, meat extract mixtures, meat mixtures and model game sausages with known red deer content. The highest accuracy was obtained when the calibration mixture was similar to the analyzed sample in both the composition and concentration of the animal species of interest. High recoveries were not only obtained for raw samples but also after subjection to thermal treatment, including brewing (15 min at 75–78 °C), boiling (90 min at 100 °C) and microwave treatment (15 s, 40 s or 2 min at 650 W). The red deer-specific real-time PCR assay was found to be robust with respect to small deviations in the reaction volume or the annealing temperature and the use of another real-time PCR instrument.

  • development and validation of a fallow deer dama dama specific taqman real time pcr assay for the detection of Food Adulteration
    Food Chemistry, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    The aim of the present study was to develop a real-time PCR assay for the identification and quantification of fallow deer (Dama dama) in Food to detect Food Adulteration. Despite high sequence homology among different deer species, a fallow deer-specific primer/probe system targeting a fragment of the nuclear MC1-R gene was designed. This primer/probe system did not amplify DNA from 19 other animals and 50 edible plant species. Moderate cross-reactivity was observed for sika deer, red deer, roe deer, reindeer and wild boar. The LOD and LOQ of the real-time PCR assay were 0.1% and 0.4%, respectively. To validate the assay, DNA mixtures, meat extract mixtures, meat mixtures and model game sausages were analyzed. Satisfactory quantitative results were obtained when the calibration mixture was similar to the analyzed sample in both the composition and concentration of the animal species of interest.

  • a novel reference real time pcr assay for the relative quantification of game meat species in raw and heat processed Food
    Food Control, 2016
    Co-Authors: Barbara Druml, Rupert Hochegger, Maria Kaltenbrunner, Margit Cichnamarkl
    Abstract:

    Abstract In order to increase their profit, Food producers may be tempted to replace expensive meat species by cheaper ones. Due to their selectivity and sensitivity, PCR based methods are frequently applied to identify and quantify meat species to detect Food Adulteration. Quantification of meat species in highly processed Food products is, however, a difficult task. When we used a previously published reference system to relatively quantify the deer content in heat treated Foodstuffs, we obtained recoveries that were substantially >100%. In the present study we aimed to improve the applicability of the reference system, targeting a 97 bp fragment of the myostatin gene, to heat treated products by reducing the length of the target sequence. The novel reference system, targeting a 70 bp fragment of the myostatin gene, was found to amplify the target region in 27 mammals and poultry species. Meat mixtures as well as raw and heat treated model sausages were analysed to demonstrate the applicability of the novel reference system for relatively quantifying the game meat content in processed Food products. With the novel reference system, the bias introduced by heat treatment was largely eliminated.

Maria Kaltenbrunner - One of the best experts on this subject based on the ideXlab platform.

  • Tetraplex real-time PCR assay for the simultaneous identification and quantification of roe deer, red deer, fallow deer and sika deer for deer meat authentication.
    Food Chemistry, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichna-markl
    Abstract:

    Abstract Analytical methods are needed for the identification and quantification of meat species to detect Food Adulteration. Since game meat is more expensive than meat from domesticated animal species, it is a potential target for Adulteration. We present a tetraplex real-time PCR assay that allows the simultaneous determination of the content of roe deer, red deer, fallow deer and sika deer. The tetraplex assay showed only moderate cross-reactivity with closely related species. After optimization the tetraplex assay had a limit of detection of 0.1% (w/w) and a limit of quantification of 0.5% (w/w) for each of the four deer species. The tetraplex assay was found to be robust, slight modifications of the experimental setup did not lower its performance. Recoveries obtained by analyzing DNA mixtures and DNA isolates from model game sausages were similar to those obtained with the singleplex assays.

  • red deer cervus elaphus specific real time pcr assay for the detection of Food Adulteration
    Food Control, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    Abstract We present a red deer-specific real-time PCR assay which, combined with a reference real-time PCR assay published previously, allows the quantification of the red deer content in Food products. Thus, it can be applied to detect Food Adulteration. The primer/probe system of the red deer-specific real-time PCR assay amplifies a 87 bp long fragment of the protein kinase C iota gene. To eliminate cross-reactivity with closely related species, the forward primer was designed to contain one deliberate base mismatch adjacent to one red deer-specific base. The red deer-specific real-time PCR assay did not show cross-reactivity with 23 animal and 50 plant species tested. LOD and LOQ, determined by analyzing a serially diluted DNA extract containing 1% (w/w) red deer DNA in pig DNA, were 0.05% and 0.4%, respectively. The accuracy was validated by analyzing DNA mixtures, meat extract mixtures, meat mixtures and model game sausages with known red deer content. The highest accuracy was obtained when the calibration mixture was similar to the analyzed sample in both the composition and concentration of the animal species of interest. High recoveries were not only obtained for raw samples but also after subjection to thermal treatment, including brewing (15 min at 75–78 °C), boiling (90 min at 100 °C) and microwave treatment (15 s, 40 s or 2 min at 650 W). The red deer-specific real-time PCR assay was found to be robust with respect to small deviations in the reaction volume or the annealing temperature and the use of another real-time PCR instrument.

  • development and validation of a fallow deer dama dama specific taqman real time pcr assay for the detection of Food Adulteration
    Food Chemistry, 2018
    Co-Authors: Maria Kaltenbrunner, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    The aim of the present study was to develop a real-time PCR assay for the identification and quantification of fallow deer (Dama dama) in Food to detect Food Adulteration. Despite high sequence homology among different deer species, a fallow deer-specific primer/probe system targeting a fragment of the nuclear MC1-R gene was designed. This primer/probe system did not amplify DNA from 19 other animals and 50 edible plant species. Moderate cross-reactivity was observed for sika deer, red deer, roe deer, reindeer and wild boar. The LOD and LOQ of the real-time PCR assay were 0.1% and 0.4%, respectively. To validate the assay, DNA mixtures, meat extract mixtures, meat mixtures and model game sausages were analyzed. Satisfactory quantitative results were obtained when the calibration mixture was similar to the analyzed sample in both the composition and concentration of the animal species of interest.

  • a novel reference real time pcr assay for the relative quantification of game meat species in raw and heat processed Food
    Food Control, 2016
    Co-Authors: Barbara Druml, Rupert Hochegger, Maria Kaltenbrunner, Margit Cichnamarkl
    Abstract:

    Abstract In order to increase their profit, Food producers may be tempted to replace expensive meat species by cheaper ones. Due to their selectivity and sensitivity, PCR based methods are frequently applied to identify and quantify meat species to detect Food Adulteration. Quantification of meat species in highly processed Food products is, however, a difficult task. When we used a previously published reference system to relatively quantify the deer content in heat treated Foodstuffs, we obtained recoveries that were substantially >100%. In the present study we aimed to improve the applicability of the reference system, targeting a 97 bp fragment of the myostatin gene, to heat treated products by reducing the length of the target sequence. The novel reference system, targeting a 70 bp fragment of the myostatin gene, was found to amplify the target region in 27 mammals and poultry species. Meat mixtures as well as raw and heat treated model sausages were analysed to demonstrate the applicability of the novel reference system for relatively quantifying the game meat content in processed Food products. With the novel reference system, the bias introduced by heat treatment was largely eliminated.

Barbara Druml - One of the best experts on this subject based on the ideXlab platform.

  • real time pcr assay for the detection and quantification of roe deer to detect Food Adulteration interlaboratory validation involving laboratories in austria germany and switzerland
    Foods, 2021
    Co-Authors: Barbara Druml, Margit Cichnamarkl, Steffen Uhlig, Kirsten Simon, Kirstin Frost, Karina Hettwer, Rupert Hochegger
    Abstract:

    Game meat products are particularly prone to be adulterated by replacing game meat with cheaper meat species. Recently, we have presented a real-time polymerase chain reaction (PCR) assay for the identification and quantification of roe deer in Food. Quantification of the roe deer content in % (w/w) was achieved relatively by subjecting the DNA isolates to a reference real-time PCR assay in addition to the real-time PCR assay for roe deer. Aiming at harmonizing analytical methods for Food authentication across EU Member States, the real-time PCR assay for roe deer has been tested in an interlaboratory ring trial including 14 laboratories from Austria, Germany, and Switzerland. Participating laboratories obtained aliquots of DNA isolates from a meat mixture containing 24.8% (w/w) roe deer in pork, roe deer meat, and 12 meat samples whose roe deer content was not disclosed. Performance characteristics included amplification efficiency, level of detection (LOD95%), repeatability, reproducibility, and accuracy of quantitative results. With a relative reproducibility standard deviation ranging from 13.35 to 25.08% (after outlier removal) and recoveries ranging from 84.4 to 114.3%, the real-time PCR assay was found to be applicable for the detection and quantification of roe deer in raw meat samples to detect Food Adulteration.

  • a novel reference real time pcr assay for the relative quantification of game meat species in raw and heat processed Food
    Food Control, 2016
    Co-Authors: Barbara Druml, Rupert Hochegger, Maria Kaltenbrunner, Margit Cichnamarkl
    Abstract:

    Abstract In order to increase their profit, Food producers may be tempted to replace expensive meat species by cheaper ones. Due to their selectivity and sensitivity, PCR based methods are frequently applied to identify and quantify meat species to detect Food Adulteration. Quantification of meat species in highly processed Food products is, however, a difficult task. When we used a previously published reference system to relatively quantify the deer content in heat treated Foodstuffs, we obtained recoveries that were substantially >100%. In the present study we aimed to improve the applicability of the reference system, targeting a 97 bp fragment of the myostatin gene, to heat treated products by reducing the length of the target sequence. The novel reference system, targeting a 70 bp fragment of the myostatin gene, was found to amplify the target region in 27 mammals and poultry species. Meat mixtures as well as raw and heat treated model sausages were analysed to demonstrate the applicability of the novel reference system for relatively quantifying the game meat content in processed Food products. With the novel reference system, the bias introduced by heat treatment was largely eliminated.

  • Duplex real-time PCR assay for the simultaneous determination of the roe deer (Capreolus capreolus) and deer (sum of fallow deer, red deer and sika deer) content in game meat products
    Food Control, 2015
    Co-Authors: Barbara Druml, Rupert Hochegger, Margit Cichna-markl
    Abstract:

    Due to the high price of game meat, Food producers may be tempted to adulterate their products with cheaper meat. This paper presents a duplex real-time PCR assay which allows the simultaneous determination of the content of roe deer (Capreolus capreolus) and deer* (the sum of fallow deer (Dama dama), red deer (Cervus elaphus) and sika deer (Cervus nippon)) in Food products to detect Food Adulteration. Relative quantification is carried out by using a reference (“all meat”) PCR assay based on the myostatin gene. The quantification approach was validated by analyzing binary meat mixtures with pork, “all game” meat mixtures containing each of the four game species in pork and a model game sausage. Compared to singleplex assays the duplex assay is time and cost saving. Thus, it is highly applicable to routine analysis in order to verify the authenticity of game meat products.

  • authenticity control of game meat products a single method to detect and quantify Adulteration of fallow deer dama dama red deer cervus elaphus and sika deer cervus nippon by real time pcr
    Food Chemistry, 2015
    Co-Authors: Barbara Druml, Stephanie Grandits, Walter Mayer, Rupert Hochegger, Margit Cichnamarkl
    Abstract:

    This contribution presents a single real-time PCR assay allowing the determination of the deer content (the sum of fallow deer (Dama dama), red deer (Cervus elaphus) and sika deer (Cervus nippon)) in meat products to detect Food Adulteration. The PCR assay does not show cross-reactivity with 20 animal species and 43 botanical species potentially contained in game meat products. The limit of quantification is 0.5% for fallow deer and red deer and 0.1% for sika deer. The deer content in meat products is determined by relating the concentration obtained with the deer PCR assay to that obtained with a reference system which amplifies mammals and poultry DNA. The analysis of binary meat mixtures with pork, a meat mixture containing equal amounts of fallow deer, red deer and sika deer in pork and a model game sausage showed that the quantification approach is very accurate (systematic error generally <25%).

Margit Cichna-markl - One of the best experts on this subject based on the ideXlab platform.