Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products

Domenico, Marco Di; Giuseppe, M. Di; Rodriguez, Jason D.; Cammà, Cesare

doi:10.3168/jds.2016-11695

Cited by 76 publications

(46 citation statements)

References 29 publications

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“…The LOD results differed according to different types of sample. The LOD for milks and dairy products were more sensitive than the previous study (Zhang et al, 2007;Di Domenico et al, 2017). However, the sensitivity for sour soup and koumiss were lower than corresponding data for cow milk, yogurt, and mare milk.…”

Section: Sensitivity In the Triplex Real-time Pcr Amplification For Mcontrasting

confidence: 58%

Simultaneous identification of bovine and equine DNA in milks and dairy products inferred from triplex TaqMan real-time PCR technique

Guo¹,

Qian²,

Guo³

et al. 2018

Journal of Dairy Science

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Koumiss is a popular dairy product in many lands, traditionally prepared from mare milk with spontaneous fermentation. Mare milk and its fermented derivates are more expensive than cow milk and its fermented derivates, and the possibility exists for producers and dealers to adulterate equine products with bovine items. In this work, we described the development of a triplex real-time PCR based on species-specific TaqMan probes for identification of bovine and equine DNA in milks and dairy products. In addition, a novel designed endogenous control was simultaneously amplified to eliminate possible false negatives. With this methodology, bovine and equine DNA were specifically identified by employing developed primers and probes. The limits of detection of this method were 0.001 ng for cow milk, yogurt, and mare milk, and 0.005 ng for sour soup and koumiss, respectively. In addition, the triplex real-time PCR assay for authentication of animal-derived products was effectively validated using binary DNA and milk mixtures, exhibiting well in terms of specificity, sensitivity, and reproducibility. In short, the triplex PCR assay was verified to be a time-saving and money-saving technique for the identification of bovine and equine DNA in milks and dairy products.

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Section: Sensitivity In the Triplex Real-time Pcr Amplification For Mcontrasting

confidence: 58%

Simultaneous identification of bovine and equine DNA in milks and dairy products inferred from triplex TaqMan real-time PCR technique

Guo¹,

Qian²,

Guo³

et al. 2018

Journal of Dairy Science

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“…These are based on antigen-antibody interactions including enzyme-linked immunosorbent assay (ELISA) [37,38], dipstick tests [39], and lateral flow devices (LFD) [40]. Furthermore, polymerase chain reaction (PCR) [41,42] and liquid chromatography-mass spectrometry (LC-MS) [26] platforms are mostly explored alternatives of antibody-based assays. Research efforts are directed for highly specific, sensitive, and rapid detection of allergens from the processed milk products [43][44][45].…”

Section: Analytical Standard Methods For the Milk Allergens Detectionmentioning

confidence: 99%

Nano-Biosensing Platforms for Detection of Cow’s Milk Allergens: An Overview

Nehra

Lettieri

Dilbaghi

et al. 2019

Sensors

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Among prevalent food allergies, cow milk allergy (CMA) is most common and may persist throughout the life. The allergic individuals are exposed to a constant threat due to milk proteins’ presence in uncounted food products like yogurt, cheese, and bakery items. The problem can be more severe due to cross-reactivity of the milk allergens in the food products due to homologous milk proteins of diverse species. This problem can be overcome by proper and reliable food labeling in order to ensure the life quality of allergic persons. Therefore, highly sensitive and accurate analytical techniques should be developed to detect the food allergens. Here, significant research advances in biosensors (specifically immunosensors and aptasensors) are reviewed for detection of the milk allergens. Different allergic proteins of cow milk are described here along with the analytical standard methods for their detection. Additionally, the commercial status of biosensors is also discussed in comparison to conventional techniques like enzyme-linked immunosorbent assay (ELISA). The development of novel biosensing mechanisms/kits for milk allergens detection is imperative from the perspective of enforcement of labeling regulations and directives keeping in view the sensitive individuals.

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“…Commission Regulation 273/2008 set isoelectric focusing of γcaseins after plasminolysis (IEF) as the reference method for the detection of cows' milk and caseinate in cheeses from ewes 'milk, goats' milk or buffalos' milk or mixtures of ewes', goats' and buffalos' milk. This is a qualitative method which although is sensitive and accurate for the detection of cow milk in mixes it has several disadvantages which include that it is not a high-throughput method, it is not quantitative, the analysis is time consuming, it cannot discriminate goat-sheep mixtures, interpretation of the IEF profile can be equivocal and is not applicable to products made of soy milk because some weak interfering bands have been observed (Addeo et al, 1990;Mayer, Heidler & Rockenbauer, 1997;Di Domenico et al, 2016). In parallel, several methods based on different techniques have been introduced in order to detect cow milk in goat milk of dairy products such as electrophoresis (Mayer, Burger & Kaar, 2012;Molina, Martin-Alvarez & Ramos, 1999), immunochemistry (Hurley, Ireland, Coleman & Williams, 2004;, chromatography (De Noni, Tirelli & Masotti, 1996;Ferreira & Caç ote, 2003;Mayer, 2005) and mass spectrometry (Cozzolino, Passalacqua, Salemi & Garozzo, 2002).…”

Section: Introductionmentioning

confidence: 99%

Milk Adulteration: Detection of Bovine Milk in Caprine Dairy Products by Real Time PCR

Tsakali¹,

Agkastra²,

Koliaki³

et al. 2019

JFR

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Milk adulteration is an international social problem. Consumption of adulterated milk may cause serious health problems and a great concern of the food industry has been raised. In this study, a method based on the polymerase chain reaction (PCR) principle was validated for detecting cow’s milk in goat's dairy products. A total of 40 goat's dairy products commonly consumed in Greece, were tested. Various concentrations, from 0.01 to 90%, of cows’ milk in goats’ milk samples were prepared for DNA extraction and further PCR analysis. Selection of highly polymorphic regions within the cow and goat mitochondrial D-loops, showing low homology between the two species, allowed to choose specific primer pairs for detection of cow and goat DNA. After electrophoresis, cow DNA was characterised by the fragment of the size of 300 bp, goat DNA by the fragment of 444 bp. The detection limit of the PCR method was 0.01% while sensitivity and specificity of the method were both 100%. Goat dairy products samples were tested for the presence of cow DNA. Thirty six out of forty (90%) that were tested, were found to produce cow-specific PCR product in addition to goat PCR product while only two samples gave goat-specific product only. The results are disappointing in terms of the food labelling honesty but on the other hand PCR is again a quickly, easy and reliable method that could be used for extended adulteration screening.

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Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products

Cited by 76 publications

References 29 publications

Simultaneous identification of bovine and equine DNA in milks and dairy products inferred from triplex TaqMan real-time PCR technique

Simultaneous identification of bovine and equine DNA in milks and dairy products inferred from triplex TaqMan real-time PCR technique

Nano-Biosensing Platforms for Detection of Cow’s Milk Allergens: An Overview

Milk Adulteration: Detection of Bovine Milk in Caprine Dairy Products by Real Time PCR

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